Note : Les descriptions sont présentées dans la langue officielle dans laquelle elles ont été soumises.
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PYRIDOQUINOXALINE ANTIVIRALS
FIELD OF THE INVENTION
The present invention provides pyridoquinoxalines that are useful as antiviral
agents. More specifically, it provides compounds of formula I described herein
below
against herpesviruses.
BACKGROUND OF THE ll~TVENTION
The herpesviruses comprise a large family of double stranded DNA viruses.
IO They are also a source of the most common viral illnesses in man. Eight of
the herpes
viruses, herpes simplex virus types l and 2 (HSV-1 and HSV-2), varicella
zoster virus
(VZV), human cytomegalovirus (HCMV), Epstein-Barn virus (EBV), and human
herpes viruses 6, 7, and 8 (HHV-6, HHV-7, and HHV-8), have been shown to
infect
humans.
HSV-l and HSV-2 cause herpetic lesions on the lips and genitals, respectively.
They also occasionally cause infections of the eye and encephalitis. HCMV
causes
birth defects in infants and a variety of diseases in immunocompromised
patients such
as retinitis, pneumonia, and gastrointestinal disease. VZV is the causitive
agent of
chicken pox and shingles. EBV causes infectious mononucleosis. It can also
cause
lymphomas in immunocompromised patients and has been associated with Burkitt's
lymphoma, nasopharyngeal carcinoma, and Hodgkins disease. HHV-6 is the
causative agent of roseola and may be associated with multiple sclerosis and
chronic
fatigue syndrome. HHV-7 disease association is unclear, but it may be involved
in
some cases of roseola. HHV-8 has been associated with Karposi's sarcoma, body
cavity based lymphomas, and multiple myeloma.
Infection by reactivation of herpesviruses is associated with several
cardiovascular diseases or conditions in the host such as atherosclerosis and
restenosis
resulting in inflammation of coronary vessel walls. It is thought that in many
patients
suffering from restenosis following coronary atherectomy viral infection
particularly
by CMV plays an important role in the proliferation of the disease.
Atherosclerosis is
believed to be be associated with the overall infectious disease burden in the
host and
particularly by the herpesviruses such as HSV, CMV, and EBV.
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Infection in the animal population (livestock and companion) by strains of
herpesviruses is endemic including cattle (Bovine herspesvirus 1-5, BHV),
sheep
(Ovine herpesvirus 1 and 2), dog (Canine herpesvirus 1), horse (Equine
herpesvirus 1-
S, EHV), cat (Feline herpesvirus 1, FHV), swine (pseudorabies virus, PRV), and
many
species of fowl. In the case of bovine herpesvirus infection, animals may
suffer from
ocular, respiratory, or digestive disorders. Pseudorabies is an extremely
contagious
viral pathogen infecting several species such as cattle, horses, dogs, cats,
sheep, and
goats leading to rapid death. The virus is benign in adult swine, however, it
remains
contagious and leads to high mortality in pigs under three weeks. Infection of
horses
by equine herpesvirus may lead to neurological syndromes, respiratory disease,
and
neonatal disease. Herpesvirus infection in cats leads to the disease known as
feline
viral rhinotracheitis (FVR) which is characterized by rhinitis, tracheitis,
laryngitis, and
conjunctivitis.
It has been surprisingly discovered that compounds of the present invention
demonstrate greatly enhanced oral bioavailability and improved selectivity for
the
viral targets.
INFORMATION DISCLOSURE
U.S. Patent No. 5,792,774 discloses specific quinoline derivatives that are
alleged to have therapeutic utility via inhibition of Phosphodiesterase IV
esterase
and/or Tumor Necrosis factor activity.
PCTlUS01/16494 discloses heterocycle carboxamides as antiviral agents.
Despite the above teachings, there still exists a need in the art for
compounds
that have enhanced oral bioavailability and improved selectivity for the viral
targets.
SUMMARY OF THE INVENTION
The present invention provides a compound of formula I,
A compound of formula I
0 0
H~N ~ I I NH W I R1
CH3 ' N'
,N\ J
R
O
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or a pharmaceutically acceptable salt thereof
wherein Rl is F or Cl;
R2 is Cl_4alkyl, optionally substituted by OH or OCl_4alkyl;
R3 is aryl or heteroaryl, optionally substituted by one to three Cl_Zalkyl,
OH,
OCl_aalkyl or CN;
aryl is a phenyl or benzyl radical optionally fused to a benzene ring; and
heteroatyl is a 5- or 6-membered aromatic ring having at least one heteroatom
selected
from the group consisting of O, S and N(X) wherein X is absent or H, wherein
heteroaryl is optionally fused to a benzene ring.
The present invention further provides:
a pharmaceutical composition comprising a compound of formula I or a
pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable
carrier;
a method of treating or preventing a herpesviral infection comprising
administering to a mammal in need of such treatment, a compound of formula I
or a
pharmaceutically acceptable salt thereof, wherein the method is administered
orally,
parenterally, topically, rectally, nasally, sublingually or transdermally;
a method for the treatment of atherosclerosis and restenosis comprising
administering to a mammal in need of such treatment, a compound of formula I
or a
pharmaceutically acceptable salt thereof, wherein the method is administered
orally,
parenterally, topically, rectally, nasally, sublingually or transdermally;
a method for the treatment of herpesviral infections comprising administering
a composition comprising a pharmaceutically effective amount of the compound
of
formula I and at least one other antiviral agent;
a method for the treatment of atherosclerosis and restenosis comprising
administering a composition comprising a pharmaceutically effective amount of
the
compound of formula I and at least one other antiviral agent;
a pharmaceutically acceptable salt thereof for use in medical treatment;
a use of a compound of formula I or a pharmaceutically acceptable salt thereof
to prepare a medicament for treating or preventing a herpesviral infection in
a
mammal; and
a method for inhibiting a viral DNA polymerase, comprising contacting (i~a
vitro or ih vivo) the polymerase with an effective inhibitory amount of a
compound of
formula I, or a pharmaceutically acceptable salt thereof.
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The invention also provides novel intermediates and processes disclosed
herein that are useful for preparing compounds of formula I.
DETAILED DESCRIPTION OF THE INVENTION
The following definitions are used, unless otherwise described. Alkyl denotes
both straight and branched groups; but reference to an individual radical such
as
"propyl" embraces only the straight chain radical, a branched chain isomer
such as
"isopropyl" being specifically referred to.
The carbon atom content of various hydrocarbon-containing moieties is
1o indicated by a prefix designating the minimum and maximum number of carbon
atoms in the moiety, i.e., the prefix Ci ~ indicates a moiety of the integer
"i" to the
integer "j" carbon atoms, inclusive. Thus, fox example, (Cl_4)alkyl refers to
alkyl of
one to four carbon atoms, inclusive, or methyl, ethyl, propyl, isopropyl and
butyl,
straight and branched forms thereof. Representative heteroaryls include
pyrazinyl;
15 furanyl; thienyl; pyridyl; pyrimidinyl; isoxazolyl; isothiazolyl; oxazolyl;
thiazolyl;
pyrazolyl; furazanyl; pyrrolyl; pyrazolyl; triazolyl; 1,2,4-thiadiazolyl;
pyrazinyl;
pyridazinyl; quinoxalinyl; phthalazinyl; 1 (2H)-phthalazinonyl;
imidazopyridinyl;
imidazothiazolyl; benzofurazanyl indolyl; azaindolyl; benzimidazolyl;
benzothienyl;
quinolinyl; imidazolyl; thienopyridyl; quinazolinyl; thienopyrimidyl;
pyrrolopyridyl;
20 imidazopyridyl; isoquinolinyl; benzoazaindolyl; azabenzimidazolyl; 1,2,4-
triazinyl;
benzothiazolyl; furopyridyl and the like.
Mammal denotes human and animals. Animals specifically refers to food
animals or companion animals.
The term "antiviral agent" refers to an antiviral drug other than a compound
of
25 formula I. Specifically, they refer to Acyclovir, Penciclovir,
Famiciclovir,
Valaciclovir, Ganciclovir, Valganciclovir, Foscarnet, and Cidofovir. Such
antiviral
agents can be either obtained commercially or be prepared according to the
references
cited in PHYSICIANS' DESK REFERENCE, the 54~' Edition (2000) and the US
FDA's Orange book.
30 Compounds of the invention may have one or more chiral centers and be
isolated in optically active and racemic forms. Some compounds may exhibit
polymorphism. It is to be understood that the present invention encompasses
any
racemic, optically-active, polymorphic, tautomeric, or stereoisomeric form, or
mixture
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thereof, of a compound of the invention, which possesses the useful properties
described herein, it being well known in the art how to prepare optically
active forms
(for example, by resolution of the racemic form by recrystallization
techniques, by
synthesis from optically-active starting materials, by chiral synthesis, or by
chromatographic separation using a chiral stationary phase) and how to
determine
antiviral activity using the standard tests described herein, or using other
similar tests
which are well known in the art.
The compounds of the present invention are generally named according to the
IUPAC or CAS nomenclature system. Abbreviations which are well known to one of
ordinary skill in the art may be used (e.g. "Ph" for phenyl, 'Me" for methyl,
"Et" for
ethyl, "h" for hour or hours and "rt" for room temperature, "N" for nitrogen
atom, "O"
for oxygen atom, and "S" for sulfur atom).
Specifically, Rl is chloro.
Specifically, Rl is fluoro.
Specifically, R2 is methyl.
Specifically, R2 is ethyl.
Specifically, RZ is ethyl substituted by OH.
Specially, R3 is phenyl.
Specifically, R3 is phenyl substituted by one or two OH or OCH3.
Specifically, R3 is a 5-membered aromatic ring having at least one heteroatom
selected from the group consisting of O, S and N(X) wherein X is absent or H.
Specifically, R3 is furyl, thienyl or thiazol, wherein R3 is optionally fused
to a
benzene ring.
Specifically, R3 is optionally substituted by one to two methyl, OH, OCH3 or
CN.
Specifically, R3 is 1-benzofuran-2-yl, 3-furyl, 2-furyl, 3-thienyl, 5-methyl-2-
furyl, 2,5-dimethyl-3-furyl, 2-thienyl, 1-benzothien-3-yl, 5-cyanothien-2-yl,
or 1,3-
thiazol-2y1.
Specifically, R3 is a 6-membered aromatic ring having at least one heteroatom
selected from the group consisting of O, S and N.
Specifically, R3 is pyridinyl, pyrimidinyl, or pyrazinyl, wherein wherein R3
is
optionally fused to a benzene ring.
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Specifically, R3 is optionally substituted by one to two methyl, OH, OCH3 or
CN.
Specifically, R3 is pyridin-2-yl, 6-methylpyridin-2-yl, pyridin-3-yl, quinolin-
2-
yl, or pyrimidin-2-yl.
Specifically, compounds of formula I includes enantionmers of formula IA:
o
NH W I Ri
HO CH3 ~N'
,N\ J
R ~2
O
IA.
Specifically, compounds of formula I includes enantionmers of formula IB:
0 0
3
NH
HO CH3 N J R
/N' J
R ~2
O
IB.
Examples of the present invention are:
(1) N-(4-chlorobenzyl)-9-{[(2-hydroxy-2-phenylethyl) (methyl)amino]methyl}-1-
methyl-2,7-dioxo-2, 3-dihydro-1 H,7H-pyrido [ 1,2, 3-de] quinoxaline-6-
carboxamide,
(2) rac N-(4-chlorobenzyl)-9-{ [[2-hydroxy-2-(4-hydroxyphenyl)ethyl]
(methyl)amino]methyl }-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[ 1,2,3-
de]quinoxaline-6-carboxarnide,
(3) rac 9-{[[2-(1-benzofuran-2-yl)-2-hydroxyethyl](methyl)amino]methyl}-N-(4-
chlorobenzyl)-1-methyl-2,7-dioxo-2, 3-dihydro-1 H,7H-pyrido [ 1,2, 3-de]
quinoxaline-6-
carboxamide,
(4) rac N-(4-chlorobenzyl)-9-{ [[2-hydroxy-2-(5-methyl-2-
furyl)ethyl](methyl)amino]methyl}-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-
pyrido [ 1,2, 3-de] quinoxaline-6-carboxamide,
(5) N-(4-chlorobenzyl)-9-{ [[(2R)-2-(2-furyl)-2-hydroxyethyl]
(methyl)amino]methyl}-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[1,2,3-
de] quinoxaline-6-carboxamide,
-6-
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(6) N-(4-chlorobenzyl)-9-{ [[(2S)-2-hydroxy-2-phenylethyl]
(methyl)amino]methyl }-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[ 1,2,3-
de]quinoxaline-6-carboxamide,
(7) N-(4-chlorobenzyl)-9-{ [[(2S)-2-hydroxy-2-pyridin-3-ylethyl]
(methyl)amino]methyl}-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[1,2,3-
de]quinoxaline-6-carboxamide,
(8) rac 9-{ [[2-(1-benzothien-3-yl)-2-hydroxyethyl](methyl)amino]methyl}-N-(4-
chlorobenzyl)-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[ 1,2,3-
de]quinoxaline-6-
carboxamide,
(9) rac N-(4-chlorobenzyl)-9-{ [(2-hydroxy-2-quinolin-2-ylethyl)
(methyl)amino]methyl }-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido [ 1,2,3-
de] quinoxaline-6-carboxamide,
(10) N-(4-chlorobenzyl)-9-{[[(2R)-2-hydroxy-2-pyrazin-2-ylethyl]
(methyl)amino]methyl }-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[ 1,2,3-
de]quinoxaline-6-carboxamide,
(11) rac 9-{[[2-(1-benzofuran-2-yl)-2-hydroxyethyl](methyl)amino]methyl}-N-(4-
fluorobenzyl)-1-methyl-2,7-dioxo-2, 3-dihydro-1 H,7H-pyrido [ 1,2, 3-de]
quinoxaline-6-
carboxamide,
(12) 9-{[[(2R)-2-(1-benzofuran-2-yl)-2-hydroxyethyl](methyl)amino]methyl}-N-
(4-chlorobenzyl)-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[1,2,3-
de]quinoxaline-6-carboxamide, or
(13) 9-{[[(2R)-2-(1-benzofuran-2-yl)-2-hydroxyethyl](methyl)amino]methyl}-N-
(4-fluorobenzyl)-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[ 1,2,3-
de]quinoxaline-
6-carboxamide.
Other examples of the present invention are:
(1) rac N-(4-chlorobenzyl)-9-{ [(2-hydroxy-2-
phenylethyl)(methyl)amino]methyl}-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-
pyrido [ 1,2,3-de] quinoxaline-6-carboxamide,
(2) rac N-(4-chlorobenzyl)-9-{ [[2-(3-furyl)-2-
hydroxyethyl](methyl)amino]methyl}-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-
pyrido[ 1,2,3-de]quinoxaline-6-carboxamide,
_7_
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(3) rac N-(4-chlorobenzyl)-9-{ [[2-(2-furyl)-2-
hydroxyethyl](methyl)amino]methyl}-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-
pyrido[1,2,3-de]quinoxaline-6-carboxamide,
(4) rac N-(4-chlorobenzyl)-9-{ [[2-(2,5-dimethyl-3-furyl)-2-
hydroxyethyl](methyl)amino]methyl}-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-
pyrido[1,2,3-de]quinoxaline-6-carboxamide,
(5) rac N-(4-chlorobenzyl)-9-{ [[2-hydroxy-2-(6-methylpyridin-2-
yl)ethyl] (methyl)amino]methyl }-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[
1,2,3-
de] quinoxaline-6-carboxamide,
l0 (6) N-(4-chlorobenzyl)-9-{ [[(2R)-2-hydroxy-2-pyridin-2-
ylethyl](methyl)amino]methyl}-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-
pyrido[1,2,3-
de] quinoxaline-6-carboxamide,
(7) rac N-(4-chlorobenzyl)-9-{ [[2-hydroxy-2-(4-hydroxy-3-
methoxyphenyl)ethyl] (methyl)amino]methyl }-1-methyl-2,7-dioxo-2,3-dihydro-
1 H,7H-pyrido [ 1,2, 3-de] quinoxaline-6-carboxamide,
(8) N-(4-chlorobenzyl)-9-{ [[(2S)-2-hydroxy-2-thien-3-
ylethyl] (methyl)amino]methyl }-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[
1,2,3-
de]quinoxaline-6-carboxamide,
(9) rac N-(4-chlorobenzyl)-9-{ [(2-hydroxy-3-
phenylpropyl)(methyl)amino]methyl}-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-
pyrido[1,2,3-de]quinoxaline-6-carboxamide, (10) racN-(4-chlorobenzyl)-9-{[[2-
hydroxy-2-(3-methoxyphenyl)ethyl] (methyl)amino]methyl }-1-methyl-2,7-dioxo-
2,3-
dihydro-1 H,7H-pyrido [ 1,2, 3-de] quinoxaline-6-carboxamide,
(11) rac N-(4-chlorobenzyl)-9-{ [(2-hydroxy-2-pyrimidin-2-
ylethyl)(methyl)amino]methyl}-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-
pyrido[1,2,3-
de] quinoxaline-6-carboxamide,
(12) rac N-( (4-chlorobenzyl)-9-{ [[2-(5-cyanothien-2-yl)-2-
hydroxyethyl] (methyl)amino]methyl }-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-
pyrido[1,2,3-de]quinoxaline-6-carboxamide,
(13) N-(4-chlorobenzyl)-9-{[[(2R)-2-hydroxy-2-(1,3-thiazol-2-
yl)ethyl] (methyl)amino]methyl}-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[
1,2,3-
de]quinoxaline-6-carboxamide,
_g_
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(14) N-(4-fluorobenzyl)-9-{[[(2R)-2-(2-fmyl)-2-
hydroxyethyl] (methyl)amino]methyl }-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-
pyrido[1,2,3-de]quinoxaline-6-carboxamide,
(15) N-(4-fluorobenzyl)-9-{[[(2R)-2-hydroxy-2-pyridin-2-
ylethyl](methyl)amino]methyl}-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-
pyrido[1,2,3-
de] quinoxaline-6-carboxamide,
(16) N-(4-fluorobenzyl)-9-{ [[(2S)-2-hydroxy-2-pyridin-3-
ylethyl](methyl)amino]methyl }-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[
1,2,3-
de] quinoxaline-6-carboxamide,
(17) rac 9-{[[2-(1-benzofuran-2-yl)-2-hydroxyethyl](methyl)amino]methyl}-N-(4-
chlorobenzyl)-1-(2-hydroxyethyl)-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[ 1,2,3-
de] quinoxaline-6-carboxamide,
(18) N-(4-fluorobenzyl)-9-{[[(2R)-2-hydroxy-2-(1,3-thiazol-2-
yl)ethyl](methyl)amino]methyl }-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido [
1,2,3-
de]quinoxaline-6-carboxamide,
(19) N-(4-fluorobenzyl)-9-{ [[(2S)-2-hydroxy-2-thien-3-
ylethyl] (methyl)amino]methyl }-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[
1,2,3-
de] quinoxaline-6-carboxamide,
(20) N-(4-fluorobenzyl)-9-{ [[(2S)-2-hydroxy-2-
2o phenylethyl](methyl)amino]methyl}-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-
pyrido[ 1,2,3-de]quinoxaline-6-carboxamide,
(21) N-(4-fluorobenzyl)-9-{ [[(2R)-2-hydroxy-2-pyrazin-2-
ylethyl] (methyl)amino]methyl }-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[
1,2,3-
de] quinoxaline-6-carboxamide,
(22) N-(4-chlorobenzyl)-9-{ [[(2R)-2-(2-furyl)-2-
hydroxyethyl](methyl)amino]methyl}-1-(2-hydroxyethyl)-2,7-dioxo-2,3-dihydro-
1 H,7H-pyrido[ 1,2,3-de] quinoxaline-6-carboxamide,
(23) N-(4-fluorobenzyl)-9-{ [[(2R)-2-hydroxy-2-thien-2-
ylethyl] (methyl)amino]methyl }-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[
1,2,3-
de]quinoxaline-6-carboxamide,
(24) 9-{[[(2S)-2-(1-benzofuran-2-yl)-2-hydroxyethyl](methyl)amino]methyl}-N-(4-
chlorobenzyl)-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[1,2,3-de]quinoxaline-
6-
carboxamide, or
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(25) 9-{[[(2S)-2-(1-benzofuran-2-yl)-2-hydroxyethyl](methyl)amino]methyl}-N-(4-
fluorobenzyl)-1-methyl-2,7-dioxo-2, 3-dihydro-1 H,7H-pyrido [ 1,2, 3-de]
quinoxaline-6-
carboxamide.
Other examples of the present invention are:
(1) N-(4-chlorobenzyl)-9-{[[(2R)-2-hydroxy-2-(4-
hydroxyphenyl)ethyl] (methyl)amino]methyl }-1-methyl-2,7-dioxo-2,3-dihydro-
1H,7H-
pyrido[1,2,3-de]quinoxaline-6-carboxamide,
(2) N-(4-chlorobenzyl)-9-{ [[(2S)-2-hydroxy-2-(4-
hydroxyphenyl)ethyl] (methyl)amino]methyl }-1-methyl-2,7-dioxo-2,3-dihydro-
1H,7H-
lo pyrido[1,2,3-de]quinoxaline-6-carboxamide,
(3) N-(4-chlorobenzyl)-9-{ [[(2R)-2-hydroxy-2-(5-methyl-2-
furyl)ethyl] (methyl)amino]methyl }-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-
pyrido[1,2,3-de]quinoxaline-6-carboxamide,
(4) N-(4-chlorobenzyl)-9-{ [[(2S)-2-hydroxy-2-(5-methyl-2-
furyl)ethyl](methyl)amino]methyl}-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-
pyrido [ 1,2, 3-de] quinoxaline-6-carboxamide,
(5) 9-{[[(2R)-2-(1-benzothien-3-yl)-2-hydroxyethyl](methyl)amino]methyl}-N-(4-
chlorobenzyl)-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[ 1,2,3-
de]quinoxaline-6-
carboxamide,
(6) 9-{[[(2S)-2-(1-benzothien-3-yl)-2-hydroxyethyl](methyl)amino]methyl}-N-(4-
chlorobenzyl)-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[ 1,2,3-
de]quinoxaline-6-
carboxamide,
(7) N-(4-chlorobenzyl)-9-{ [((2R)-2-hydroxy-2-quinolin-2-
ylethyl)(methyl)amino]methyl }-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[
1,2,3-
de]quinoxaline-6-carboxamide,
(8) N-(4-chlorobenzyl)-9-{ [((2S)-2-hydroxy-2-quinolin-2-
ylethyl)(methyl)amino]methyl }-1-methyl-2,7-dioxo-2,3-dihydro-1 H,7H-pyrido[
1,2,3-
de] quinoxaline-6-carboxamide
(9) 9-{ [[(2R)-2-(1-benzofuran-2-yl)-2-hydroxyethyl](methyl)amino]methyl}-N-
(4-fluorobenzyl)-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[1,2,3-
de]quinoxaline-
6-carboxamide, or
- l0-
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(10) 9-{ [[(2S)-2-(1-benzofuran-2-yl)-2-hydroxyethyl](methyl)amino]methyl}-N-
(4-
fluorobenzyl)-1-methyl-2,7-dioxo-2, 3-dihydro-1 H,7H-pyrido [ 1,2,3-de]
quinoxaline-6-
carboxamide.
As shown in Chart A, the starting material A-0, whose preparation is
described in US patent # 6,093,732, is activated with methanesulfonyl chloride
and
treated with secondary amines to provide compounds of formula A-1, wherein R'
is
aryl or heteroaryl. Compounds of formula A-1 are alkylated with short chain
alkyl or
phenyl bromoacetates to give compounds of formula A-2, wherein R ~ is a short
chain
alkyl or phenyl. Compounds of formula A-2 are reacted with primary amines in
1o suitable solvents (e.g, methanol or THF) to yield amides of formula A-3.
Compounds
of formula A-3 are reacted with a suitable base (e.g., KOtBu) in the
appropriate
solvent (e.g., THF) to provide compounds of formula A-4. The secondary amines
which will comprise a portion of compounds of formula A-4 can be utilized as a
racemic mixture comprising a 1:1 mixture of the R- and S-optical antipodes.
Alternatively, secondary amines which will comprise a portion of compounds of
formula A-4 can be as optically enriched or single optical antipodes (either
the R- or
S- antipode dominates or is the sole constituent). The secondary amines afford
compounds which are racemates or configured as previously illustrated for
compounds of formulas IA and IB as a function of the nature of the amine
utilized.
CHART A
HO O HO O
HO ~ ~ N / R' N ~ ~ N
I H~ ---~ I H~ -->
\ N ~ CI OH CH3 \ N \ CI
F F A-1
A-0
O O O O
R I.. R ;,~ / N
CH3 ~ I N I H ~ I CI ~ "" CHa ' I N I H ~ I CI
F ~R" F ~NHR2
A_2 ~O A.3 O
O O
R'
~~H ~I NI H ~I
3 ~ CI
~N
R2
O A-4
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As shown in Chart B, the starting material B-0, whose preparation is described
in US patent # 6,093,732, is alkylated with a short chain alkyl or phenyl
bromoacetate
to give the esters of formula B-1, wherein R° can be a short chain
alkyl or phenyl.
Compounds of formula B-1 are reacted with various primary amines in suitable
solvents (e.g, methanol or THF) to yield amides of formula B-2. Compounds of
formula B-2 are reacted with KOtBu in THF to provide compounds of formula B-3.
Compounds of formula B-3 are treated with ethyl chlorofomate to give compounds
of
formula B-4. Compounds of formula B-4 are reacted with secondary amines to
yield
1o compounds of formula B-5, wherein R' is aryl or heteroaryl. The secondary
amines
which will comprise a portion of compounds of formula B-5 can be utilized as a
racemic mixture comprising a 1:1 mixture of the R- and S-optical antipodes.
Alternatively, secondary amines which will comprise a portion of compounds of
formula B-5 can be as optically enriched or single optical antipodes (either
the R- or
S- antipode dominates or is the sole constituent). The secondary amines afford
compounds which are racemates or configured as previously illustrated for
compounds of formulas IA and IB as a function of the nature of the amine
utilized.
CHART B
HO O O O
N ~ ~ N ~ N ~' N
SIN H ylCl-> J MINI H ~IC-->
F F ~O
B.0 B.1
OR°
O O O O
J ~I ~H ~I -> J ~I I H ~I ->
N CI NJ CI
F ~O R2iN ll
B-3
g-2 NHR2 O
O O O O
R'
CI ~I I H ~I -> ~N ~I I H ~I
N ~ CI OH CH3 N CI
R~N~ R2~N
O B-4 O B-5
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Pharmaceutical Salts
The compound of formula I may be used in its native form or as a salt. In
cases where forming a stable nontoxic salt is desired, administration of the
compound
as a pharmaceutically acceptable salt may be appropriate. Examples of
pharmaceutically acceptable salts are organic acid addition salts formed with
acids
which form a physiological acceptable anion, for example, tosylate,
methanesulfonate,
acetate, citrate, malonate, tartarate, succinate, benzoate, ascorbate,
ketoglutarate, and
glycerophosphate. Suitable inorganic salts may also be formed, including
hydrochloride, hydrobromide, sulfate, nitrate, bicarbonate, and carbonate
salts.
Pharmaceutically acceptable salts may be obtained using standard procedures
well known in the art, for example by reacting a compound of the invention
with a
suitable acid affording a physiologically acceptable anion.
Routes of Administration
In therapeutic use for treating, or combating, viral infections in a mammal
(i.e.
human and animals) a compound of the present invention, its pharmaceutical
compositions and other antiviral agents can be administered orally,
parenterally,
topically, rectally, transmucosally, or intestinally.
Parenteral administrations include indirect injections to generate a systemic
effect or direct injections to the afflicted area. Examples of parenteral
administrations
are subcutaneous, intravenous, intramuscular, intradermal, intrathecal,
intraocular,
intranasal, intravetricular injections or infusions techniques.
Topical administrations include the treatment of infectious areas or organs
readily accessibly by Iocal application, such as, for example, eyes, ears
including
external and middle ear infections, vaginal, open wound, skins including the
surface
skin and the underneath dermal structures, or other lower intestinal tract. It
also
includes transdermal delivery to generate a systemic effect.
The rectal administration includes the form of suppositories.
The transmucosal administration includes nasal aerosol or inhalation
applications.
The preferred routes of administration are oral and parenteral.
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Composition/Formulation
Pharmaceutical compositions of the present invention may be manufactured by
processes well known in the art, e.g., by means of conventional mixing,
dissolving,
granulation, dragee-making, levigating, emulsifying, encapsulating,
entrapping,
lyophilizing processes or spray drying.
Pharmaceutical compositions for use in accordance with the present invention
may be formulated in conventional manner using one or more physiologically
acceptable carriers comprising excipients and auxiliaries which facilitate
processing of
the active compounds into preparations which can be used pharmaceutically.
Proper
formulation is dependent upon the route of administration chosen.
For oral administration, the compounds can be formulated by combining the
active compounds with pharmaceutically acceptable carriers well known in the
art.
Such carriers enable the compounds of the invention to be formulated as
tablets, pills,
lozenges, dragees, capsules, liquids, solutions, emulsions, gels, syrups,
slurries,
suspensions and the like, for oral ingestion by a patient. A carrier can be at
least one
substance which may also function as a diluent, flavoring agent, solubilizer,
lubricant,
suspending agent, binder, tablet disintegrating agent, and encapsulating
agent.
Examples of such carriers or excipients include, but are not limited to,
magnesium
carbonate, magnesium stearate, talc, sugar, lactose, sucrose, pectin, dextrin,
mannitol,
sorbitol, starches, gelatin, cellulosic materials, low melting wax, cocoa
butter ox
powder, polymers such as polyethylene glycols and other pharmaceutical
acceptable
materials.
Dragee cores are provided with suitable coatings. For this purpose,
concentrated sugar solutions may be used which may optionally contain gum
arabic,
talc, polyvinyl pyrrolidone, carbopol gel, polyethylene glycol, and/or
titanium dioxide,
lacquer solutions, and suitable organic solvents or solvent mixtures.
Dyestuffs or
pigments may be added to the tablets or dragee coatings for identification or
to
characterize different combinations of active compound doses.
3o Pharmaceutical compositions which can be used orally include push-fit
capsules made of gelatin, as well as soft, sealed capsules made of gelatin and
a
plasticizer, such as glycerol or sorbitol. The push-fit capsules can contain
the active
ingredients in admixture with a filler such as lactose, a binder such as
starch, and/or a
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lubricant such as talc or magnesium stearate and, optionally, stabilizers. In
soft
capsules, the active compounds may be dissolved or suspended in suitable
liquids,
such as fatty oils, liquid paraffin, liquid polyethylene glycols, cremophor,
capmul,
medium or long chain mono-, di- or triglycerides. Stabilizers may be added in
these
formulations, also.
Liquid form compositions include solutions, suspensions and emulsions. For
example, there may be provided solutions of the compounds of this invention
dissolved in water and water-propylene glycol and water-polyethylene glycol
systems,
optionally containing suitable conventional coloring agents, flavoring agents,
stabilizers and thickening agents.
The compounds may also be formulated for parenteral administration, e.g., by
injection, bolus injection or continuous infusion. Formulations for parenteral
administration may be presented in unit dosage form, e.g., in ampoules or in
multi-
dose containers, with an added preservative. The compositions may take such
forms'
as suspensions, solutions or emulsions in oily or aqueous vehicles, and may
contain .
formulating materials such as suspending, stabilizing and/or dispersing
agents.
For injection, the compounds of the invention may be formulated in aqueous
solution, preferably in physiologically compatible buffers or physiological
saline
buffer. Suitable buffering agents include trisodium orthophosphate, sodium
2o bicarbonate, sodium citrate, N-methylglucamine, L(+)-lysine and L(+)-
arginine.
Parenteral administrations also include aqueous solutions of a water soluble
form, such as, without limitation, a salt, of the active compound.
Additionally,
suspensions of the active compounds may be prepared in a lipophilic vehicle.
Suitable lipophilic vehicles include fatty oils such as sesame oil, synthetic
fatty acid
esters such as ethyl oleate and triglycerides, or materials such as liposomes.
Aqueous
injection suspensions may contain substances which increase the viscosity of
the
suspension, such as sodium carboxymethyl cellulose, sorbitol, or dextran.
Optionally,
the suspension may also contain suitable stabilizers and/or agents that
increase the
solubility of the compounds to allow for the preparation of highly
concentrated
solutions.
Alternatively, the active ingredient may be in powder form for constitution
with a suitable vehicle, e.g., sterile, pyrogen-free water, before use.
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For suppository administration, the compounds may also be formulated by mixing
the
agent with a suitable non-irritating excipient which is solid at room
temperature but
liquid at rectal temperature and therefore will melt in the rectum to release
the drug.
Such materials include cocoa butter, beeswax and other glycerides.
For administration by inhalation, compounds of the present invention can be
conveniently delivered through an aerosol spray in the form of solution, dry
powder,
or suspensions. The aerosol may use a pressurized pack or a nebulizer and a
suitable
propellant. In the case of a pressurized aerosol, the dosage unit may be
controlled by
providing a valve to deliver a metered amount. .Capsules and cartridges of,
for
example, gelatin for use in an inhaler may be formulated containing a power
base such
as lactose or starch.
For topical applications, the pharmaceutical composition may be formulated in
a suitable ointment containing the active component suspended or dissolved in
one or
more carriers. CaiTiers for topical administration of the compounds of this
invention
include, but are not limited to, mineral oil, liquid petrolatum, white
petrolatum,
propylene glycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax
and water. Alternatively, the pharmaceutical compositions can be formulated in
a
suitable lotion such as suspensions, emulsion, or cream containing the active
components suspended or dissolved in one or more pharmaceutically acceptable
carriers. Suitable carriers include, but are not limited to, mineral oil,
sorbitan
monostearate, polysorbate 60, cetyl esters wax, ceteary alcohol, 2-
octyldodecanol,
benzyl alcohol and water.
For ophthalmic and otitis uses, the pharmaceutical compositions may be
formulated as micronized suspensions in isotonic, pH adjusted sterile saline,
or
preferably, as solutions in isotonic, pH adjusted sterile saline, either with
or without a
preservative such as a benzylalkonium chloride. Alternatively, for ophthalmic
uses,
the pharmaceutical compositions may be formulated in an ointment such as
petrolatum.
In addition to the formulations described previously, the compounds may also
be formulated as depot preparations. Such long acting formulations may be in
the
form of implants. A compound of this invention may be formulated for this
route of
administration with suitable polymers, hydrophobic materials, or as a sparing
soluble
derivative such as, without limitation, a sparingly soluble salt.
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Additionally, the compounds may be delivered using a sustained-release
system. Various sustained-release materials have been established and are well
known
by those skilled in the art. Sustained-release capsules may, depending on
their
chemical nature, release the compounds for 24 hours or for up to several days.
Dosage
Pharmaceutical compositions suitable for use in the present invention include
compositions wherein the active ingredients are contained in an amount
sufficient to
achieve the intended purpose, i.e., the treatment or prevention of infectious
diseases.
More specifically, a therapeutically effective amount means an amount of
compound
l0 effective to prevent, alleviate or ameliorate symptoms of disease or
prolong the
survival of the subject being treated.
The quantity of active component, that is the compound of this invention, in
the pharmaceutical composition and unit dosage form thereof may be varied or
adjusted widely depending upon the manner of administration, the potency of
the
particular compound and the desired concentration. Determination of a
therapeutically
effective amount is well within the capability of those skilled in the art.
Generally, the
quantity of active component will range between 0.5% to 90% by weight of the
composition.
Generally, an antiviral effective amount of dosage of active component will be
in the range of about 0.1 to about 400 mg/kg of body weight/day, more
preferably
about 1.0 to about 50 mglkg of body weight/day. It is to be understood that
the
dosages may vary depending upon the requirements of each subject and the
severity of
the viral infection being treated. In average, the effective amount of active
component
is about 200/mg to 800/mg and preferable 600/mg per day.
The desired dose may conveniently be presented in a single dose or as divided
doses administered at appropriate intervals, for example, as two, three, four
or more
sub-doses per day. The sub-dose itself may be further divided, e.g., into a
number of
discrete loosely spaced administrations; such as multiple inhalations from an
insufflator or by application of a plurality of drops into the eye.
Also, it is to be understood that the initial dosage administered may be
increased beyond the above upper level in order to rapidly achieve the desired
plasma
concentration. On the other hand, the initial dosage may be smaller than the
optimum
and the daily dosage may be progressively increased during the course of
treatment
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depending on the particular situation. If desired, the daily dose may also be
divided
into multiple doses for administration, e.g., two to four times per day.
In cases of local administration or selective uptake, the effective local
concentration of the drug may not be related to plasma concentration and other
procedures know in the art may be used to determine the desired dosage amount.
Combination Therapy
In combating the infective diseases caused by viruses, the compound of the
formula I can be used either individually, or in combination with other
antiviral agents
that are active against diseases caused by viruses.
Examples of such antiviral agents and the amount, route, and frequency of
administration of such active agents are listed in Table 1.
Table 1.
I
Active Agents
Used in
Combination
with a Comuound
of Formula
(ID
Active AgentLo Dose Hi Dose Route Frequency
of
Administration
Acyclovir 100 mg 800 mg Oral 2-5 times/day
Acyclovir 1% ointment 5% ointment Topical ----
Penciclovir 1% ointment 5% ointment Topical ----
Famciclovir 100 mg 500 mg Oral 1-3 times/day
Valaciclovir250 mg 1000 mg Oral 1-3 times/day
Ganciclovir 1 mg/kg 5 mg/kg IV 1-2 times/day
Ganciclovir 500 mg 1000 mg Oral 2-6 times/day
Valganciclovir200 mg/day 2000 mg/day Oral ----
Foscarnet 20 mg/kg/day120 mg/kg/dayIV ----
Cidofovir 1 mg/kg/week5 mg/kg/week IV ----
The term "Lo Dose" means the recommended lower dosage for the
combination therapy of the invention. It may be adjusted even lower depending
on the
requirements of each subject being treated and the severity of the viral
infection. The
lowest dosage possible may be 0.1 mg when combined with the compound of
formula
(II) of the present invention. The term "Hi Dose" means the recommended
highest
dosage in the combination therapy. It may be changed hereafter according to
the US
FDA standard. A specific active agent may have more than one recommended
dosage range, particularly for different routes of administration.
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For the combination therapy, the compound of formula I may be administered
concurrently or concomitantly with other antiviral agents. The term
"concurrently"
means the subject being treated takes one drug within about 5 minutes of
taking the
other drug. The term "concomitantly" means the subject being treated takes one
drug
within the same treatment period of taking the other chug. The same treatment
period
is preferably within twelve hours and up to forty-eight hours.
For the combination therapy, the compound of formula I, and one or more
other antiviral agents may be administered in the same physical form or
separately,
i.e., they may be administered in the same delivery vehicle or in different
delivery
vehicles.
Specifically, the combination therapy of the present invention is the compound
of formula I of the present invention with Acyclovir.
Specifically, the combination therapy of the present invention is the compound
of formula I of the present invention with Penciclovir.
Specifically, the combination therapy of the present invention is the compound
of formula I of the present invention with Famciclovir.
Specifically, the combination therapy of the present invention is the compound
of formula I of the present invention with Valaciclovir.
Specifically, the combination therapy of the present invention is the compound
of formula I of the present invention with Ganciclovir.
Specifically, the combination therapy of the present invention is the compound
of formula I of the present invention with Valganciclovir.
Specifically, the combination therapy of the present invention is the compound
of formula I of the present invention with Foscarnet.
Specifically, the combination therapy of the present invention is the compound
of formula I of the present invention with Cidofovir.
BIOLOGICAL DATA
While many of the compounds of the present invention have shown activity
against the CMV polymerase, these compounds may be active against the
3o cytomegalovirus by this or other mechanisms of action. Thus, the
description below
of these compounds' activity against the CMV polymerase is not meant to limit
the
present invention to a specific mechanism of action.
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The compounds of the present invention have shown activity in one or more of
the assays described below. All of these assays are indicative of a compound's
activity
and thus of its use as an anti-viral agent.
The HCMV polymerase assay is performed using a scintillation proximity
assay (SPA) as described in several references, such as N.D. Cook, et al.,
Pharmaceutical Manufacturing International, pages 49-53 (1992); K. Takeuchi,
Laboratory Practice, September issue (1992); US Patent No. 4,568,649 (1986);
which
are incorporated by reference herein. Reactions are performed in 96-well
plates. The
assay is conducted in 100 ~,l volume with 5.4 mM HEPES (pH 7.5), 11.7 mM KCl,
l0 4.5 mM MgCl2, 0.36 mg/ml BSA, and 90 nM 3H-dTTP. Assays are run with and
without CHAPS, (3-[(3-Cholamidopropyl)-dimethylammonio]-1-propane-sulfonate)
at a final concentration of 2 mM. HCMV polymerase is diluted in enzyme
dilution
buffer containing 50% glycerol, 250 mM NaCI, 10 rnM HEPES (pH 7.5), 100
~,g/rnl
BSA, and 0.01% sodium azide. The HCMV polymerase, which is expressed in
recombinant baculovirus-infected SF-9 cells and purified according to
literature
procedures, is added at 10% (or 10 ~1) of the final reaction volume, i.e., 100
~ul.
Compounds are diluted in 50% DMSO and 10 ~1 are added to each well. Control
wells contain an equivalent concentration of DMSO. Unless noted otherwise,
reactions are initiated via the addition of 6 nM biotinylated poly(dA)-
oligo(dT)
2o template/primer to reaction mixtures containing the enzyme, substrate, and
compounds of interest. Plates are incubated in a 25 °C or 37 °C
H20 bath and
terminated via the addition of 40 ~,1/reaction of 0.5 M EDTA (pH 8) per well.
Reactions are terminated within the time-frame during which substrate
incorporation
is linear and varied depending upon the enzyme and conditions used, i.e., 30
min. for
HCMV polymerase. Ten (10) ~ul of streptavidin-SPA beads (20 mg/ml in PBSllO%
glycerol) are added following termination of the reaction. Plates are
incubated 10
min. at 37 °C, then equilibrated to room temperature, and counted on a
Packard
Topcount. Linear regressions are performed and ICSO s are calculated using
computer
software.
A modified version of the above HCMV polymerase assay is performed as
described above, but with the following changes: Compounds are diluted in 100%
DMSO until final dilution into assay buffer. In the previous assay, compounds
are
diluted in 50% DMSO. 4.5 mM dithiotherotol (DTT) is added to the polymerase
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buffer. Also, a different lot of CMV polymerase is used, which appears to be
more
active resulting in a more rapid polymerase reaction.
Results of the testing of compounds of the present invention in this assay are
shown in Tables 2 below.
All results are listed as Polymerase ICSO (~M) values. In Table 2, the term
"--" refers to activity data not determined.
CMV Alpha
Pol merasePolymerase
Exam le No. IC50 (uM) IC50 (uM)
1 0.220 --
2 0.070 >5.0
0.0930 --
3 0.0086 >5.0
0.0078 --
0.0072 --
4 0.570 --
0.240 --
6 0.120 >20.0
0.0890 --
7 0.250 --
8 1.10 --
9 0.300 --
0.240 --
11 0.110 >20.0
0.0460 --
12 0.130 20.0
0.0330 --
13 0.150 >20.0
0.0760 --
14 0.190 --
0.110 >20.0
0.0860 --
16 0.0480 >20.0
0.0360 --
17 0.870 --
18 0.180 --
19 0.490 --
0.410 --
21 0.160 >10.0
0.120 --
22 0.490 --
23 0.420 --
24 1.870 --
0.930 --
26 0.880 --
27 2.080 --
28 0.690 --
29 0.810 --
1.090 --
31 2.890 --
32 0.580 --
33 0.032 4.3900
34 0.0040 -
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CMV Alpha
Polymerase Polymerase
Exam le IC50 (uM) IC50 (uM)
No.
35 0.180 --
36 <0.03 --
1.19 __
Without further elaboration, it is believed that one skilled in the art can,
using
the preceding description, practice the present invention to its fullest
extent. The
foregoing detailed description is given for clearness of understanding only,
and no
unnecessary limitations should be understood therefrom, as modifications
within the
scope of the invention may become apparent to those skilled in the art.
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EXAMPLES
Example 1 Preparation of N-(4-chlorobenzyl)-9-{ [(2-hydroxy-2-phenylethyl)
(methyl)amino]methyl}-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[1,2,3-
de] quinoxaline-6-carboxamide.
iI o 0
OH ~ ~ I I NH
'N' CI
iN
5.
Step 1 Preparation of methyl [3-{ [(4-chlorobenzyl)amino]carbonyl}-8-fluoro-
6-(morpholin-4-ylmethyl)-4-oxoquinolin-1 (4H)-yl] acetate.
To a flask containing N-(4-chlorobenzyl)-8-fluoro-4-hydroxy-6-(morpholin-4-
ylmethyl)quinoline-3-carboxamide (preparation described in US patent #
6,093,732)
(10.8 g) is added dimethylformamide (DMF) (100 mL) and potassium carbonate
(8.85
g). The resulting suspension is placed under a drying tube and stirred. To the
stirring
mixture is added methyl bromoacetate (4.8 mL). After stirring overnight, the
suspension is diluted with water (250 mL) and filtered. The precipitant is
washed with
two additional portions of water (2 x 100 mL). The washed solid is dried under
a flow
of air and then in a vacuum oven (60 °C) to yield 12.2 g of the title
compound as a tan
solid. Physical characteristics: 1H NMR (CDC13) ~ 2.5, 3.6, 3.7, 3.8, 4.6,
5.1, 7.3, 7.5,
8.2, 8.7, 10.3; MS (ESI+) for m/z 502 (M+H)+.
Step 2 Preparation of N-(4-chlorobenzyl)-8-fluoro-1-[2-(methylamino)-2
oxoethyl]-6-(morpholin-4-ylmethyl)-4-oxo-1,4-dihydroquinoline-3-carboxamide.
To a pressure tube containing methyl [3-{ [(4-chlorobenzyl)amino]carbonyl }-
8-fluoro-6-(morpholin-4-ylmethyl)-4-oxoquinolin-1(4H)-yl]acetate (0.48 g) is
added
methylamine as a 2 M solution in methanol ( 12 mL). The mixture is tightly
capped,
heated to 60°C and stirred. After 2 days, the suspension is cooled to
room temperature
and concentrated under reduced pressure. The residue is treated with
acetonitrile:methanol (1:1, 50 mL) and heated to reflux. The resulting
suspension is
cooled to room temperature and placed in a freezer. The solid is collected,
washed
with diethyl ether and hexanes, dried under a flow of air, and finally in a
vacuum oven
(60 °C) to yield 0.47 g of the title compound as a white solid.
Physical characteristics:
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1H NMR (d6-DMSO) 8 2.4, 2.7, 3.6, 4.6, 5.5, 7.4, 7.6, 8.1, 8.3, 8.8, 10.3; MS
(ESI+)
for fnlz 501 (M+H)+.
Step 3 Preparation of N-(4-chlorobenzyl)-1-methyl-9-(morpholin-4-ylmethyl)-
2,7-dioxo-2,3-dihydro-1H,7H-pyrido[1,2,3-de]quinoxaline-6-carboxamide.
To a flame-dried flask under a nitrogen atmosphere is added N-(4-
chlorobenzyl)-8-fluoro-1-[2-(methylamino)-2-oxoethyl]-6-(morpholin-4-ylmethyl)-
4-
oxo-1,4-dihydroquinoline-3-carboxamide (0.10 g) followed by THF (10 mL). The
mixture is treated with a solution of potassium tert-butoxide in THF (1 M,
0.20 mL).
After 3 hours, the resulting dark red solution is diluted with dichloromethane
and
partitioned against pH 7 aqueous phosphate buffer. The aqueous layer is
extracted
with two additional portions of dichloromethane. The combined organic layer is
washed brine, dried over sodium sulfate, filtered and concentrated under
reduced
pressure. The resulting residue is flash column chromatographed on silica
eluting with
1% to 5% methanol in dichloromethane. The product containing fractions are
combined and concentrated under reduced pressure. The resulting residue is
crystallized from acetonitrile:methanol. The solid is collected to yield 0.04
g of the
title compound as a white solid. Physical characteristics: m.p. 213-
214°C; analysis
found for C25H25C1N~O4: C 62.27; H, 5.25; N, 11.60; 1H NMR (d6-DMSO) 8 2.4,
3.4,
3.6, 4.6, 5.2, 7.4, 7.9, 8.8, 10.4; MS (ESI+) for m/z 481 (M+H)+.
Step 4 Preparation of N-(4-chlorobenzyl)-9-(chloromethyl)-1-methyl-2,7-
dioxo-2,3-dihydro-1 H,7H-pyrido [ 1,2, 3-de] quinoxaline-6-carboxamide.
To a pressure tube containing N-(4-chlorobenzyl)-1-methyl-9-(morpholin-4-
ylmethyl)-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[1,2,3-de]quinoxaline-6-
carboxamide
(0.10 g) is added chloroform (2 mL) and ethyl chloroformate (0.05 mL). The
solution
is tightly capped, heated to 60°C and stirred overnight. The resulting
suspension is
cooled to room temperature and diluted with diethyl ether. The mixture is
filtered and
the collected precipitant washed with diethyl ether and hexanes, dried under a
flow of
3o air, and finally in a vacuum oven to yield 0.09 g of the title compound as
a white
solid. Physical characteristics:1H NMR (d6-DMSO) ~ 3.4, 4.6, 5.0, 5.2, 7.4,
7.6, 8.0,
8.8, 10.3.
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Step 5 Preparation of N-(4-chlorobenzyl)-9-{ [(2-hydroxy-2-
phenylethyl)(methyl)amino]methyl }-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-
pyrido [ 1,2, 3-de] quinoxaline-6-carboxamide
To a flask containing N-(4-chlorobenzyl)-9-(chloromethyl)-1-methyl-2,7-dioxo-
2,3-
dihydro-1H,7H-pyrido[1,2,3-de]quinoxaline-6-carboxamide (0.09 g) is added a-
(methylaminomethyl)benzyl alcohol (0.05 g), diisopropylethylamine (0.05 mL)
and
DMF (2 mL). The resulting suspension is stirred at room temperature. After 4
days,
the reaction mixture is diluted with ethyl acetate (50 mL), washed with pH 7
phosphate buffer (3 x 10 mL), brine, dried over sodium sulfate, filtered, and
finally
1o concentrated under reduced pressure. The residue is flash column
chromatographed on
silica eluting with 2% to 4% methanol in dichloromethane. The product
containing
fractions are combined and concentrated under reduced pressure. The resulting
residue
is crystallized from ethyl acetate:hexanes to yield 0.04 g of the title
compound as a
white solid. Physical characteristics:1H NMR (d6-DMSO) ~ 2.3, 2.5, 3.3, 3.7,
4.6, 4.8,
5.1, 5.2, 7.2-7.5, 7.8, 8.7, 10.4; MS (ESI+) for m/z 545 (M+H)+.
EXAMPLE 2 Preparation of rac N-(4-chlorobenzyl)-9-{([2-hydroxy-2-(4-
hydroxyphenyl)ethyl] (methyl) amino)methyl } -1-methyl-2,7-dioxo-2, 3-dihydro-
1 H,7H-
2o pyrido[1,2,3-de]quinoxaline-6-carboxamide.
Ho ~ I o 0
~N ~ NH
OH ~
N CI
~N~
~O
To a flask containing N-(4-chlorobenzyl)-~9-(chloromethyl)-1-methyl-2,7-dioxo-
2,3-
dihydro-1H,7H-pyrido[1,2,3-de]quinoxaline-6-carboxamide (0.43 g) and
synephrine
(0.27 g) is added DMF ( 10 mL) and diisopropylethylamine (0.80 mL). After 3
days of
stirring at room temperature, the reaction mixture is diluted with ethyl
acetate ( 150
mL) and washed with dilute pH 4 phosphate buffer, dilute pH 7 phosphate
buffer,
brine, dried over sodium sulfate, filtered, and finally concentrated under
reduced
pressure. The residue is adsorbed onto silica gel and flash column
chromatographed
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eluting with 2% to 10% methanol in dichloromethane. The product containing
fractions are combined and concentrated under reduced pressure. The resulting
residue
is crystallized from acetonitrile and is then recrystallized from methanol -
toluene to
yield 0.14 g of the title compound as a white solid. Physical characteristics:
Mp 145-
148 °C;1H NMR (d6-DMSO) 8 10.4, 9.22, 8.73, 7.79, 7.4, 7.09, 6.67,
5.21, 4.94, 4.7,
4.57, 3.7, 3.3, 2.6-2.4, 2.23; HRMS (ESI+) m/z 561.1881 (M+H)+; Anal. Found:
C,
64.16; H, 5.12; N, 9.93.
EXAMPLE 3 Preparation of rac 9-{([2-(1-benzofuran-2-yl)-2-
l0 hydroxyethyl](methyl)amino)methyl}-N-(4-chlorobenzyl)-1-methyl-2,7-dioxo-
2,3-
dihydro-1H,7H-pyrido [ 1,2,3-de]quinoxaline-6-carboxamide.
~I I o 0
N ~I I NH ~I
OH I ~ N ~CI
~N~
[~O
To a flask containing N-(4-chlorobenzyl)-9-(chloromethyl)-1-methyl-2,7-dioxo-
2,3-
dihydro-1H,7H-pyrido[1,2,3-de]quinoxaline-6-carboxamide (0.21 g) and 1-(1-
benzofuran-2-yl)-2-(rnethylamino)ethanol (0.13 g) is added DMF (5 mL) and
diisopropylethylanune (0.5 mL,). After 3 days of stirring at room temperature,
the
reaction mixture is diluted with ethyl acetate (100 mL), washed with dilute pH
4
phosphate buffer (4 x 10 mL), brine, dried over sodium sulfate, filtered, and
finally
concentrated under reduced pressure. The residue is flash column
chromatographed on
silica eluting with 4% to 9% methanol in ethyl acetate. The product containing
fractions are combined and concentrated under reduced pressure. The resulting
residue
is crystallized from toluene to yield 0.22 g of the title compound as an off
white solid.
Physical characteristics: Mp 136-139 °C;1H NMR (d6-DMSO) S 10.4,
8.72, 7.77,
7.54, 7.4, 7.2, 7.12, 6.74, 5.55, 5.17, 4.9, 4.57, 3.7, 3.00, 2.9, 2.5, 2.30;
HRMS (ESI+)
m/z 585.1899 (M+H)+; Anal. Found: C, 65.42; H, 5.19; N, 9.59.
EXAMPLE 4 Preparation of rac N-(4-chlorobenzyl)-9-{ ([2-(3-furyl)-2-
hydroxyethyl](methyl)amino)methyl}- 1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-
pyrido [ 1,2, 3-de] quinoxaline-6-carboxamide.
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O O
i
~ ~I I NH ~~I
OH ~ N ~CI
~N~
nO
To a flask containing N-(4-chlorobenzyl)-9-(chloromethyl)-1-methyl-2,7-dioxo-
2,3-
dihydro-1H,7H-pyrido[1,2,3-de]quinoxaline-6-carboxamide (0.2I g) and 1-(3-
furyl)-
2-(methylamino)ethanol (O.IO g) is added DMF (5 mL) and diisopropylethylamine
(0.5 mL). After 3 days of stirring at room temperature, the reaction mixture
is diluted
with ethyl acetate ( 100 mL), washed with dilute pH 4 phosphate buffer (4 x 10
mL),
brine, dried over sodium sulfate, filtered, and finally concentrated under
reduced
to pressure. The residue is flash column chromatographed on silica eluting
with 4% to
9% methanol in ethyl acetate. The product containing fractions are combined
and
concentrated under reduced pressure. The resulting residue is crystallized
from
toluene to yield 0.20 g of the title compound as a white solid. Physical
characteristics:
Mp 145-148 °C;1H NMR (d6-DMSO) 8 10.4, 8.73, 7.81, 7.6, 7.4, 6.43,
5.22, 5.02,
4.75, 4.57, 3.7, 3.35, 2.7-2.5, 2.24; HRMS (ESI+) rrrJz 535.1770 (M+H)+; Anal.
Found: C, 62.63; H, 5.18; N, 10.42.
EXAMPLE 5 Preparation of sac N-(4-chlorobenzyl)-9-{([2-(2-furyl)-2-
hydroxyethyl] (methyl)amino)methyl }-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-
pyrido[1,2,3-de]quinoxaline-6-carboxamide.
0 0
~N ~I I N" ~I
OH ~ ~ N ~CI
~N~
~O
To a flask containing N-(4-chlorobenzyl)-9-(chloromethyl)-1-methyl-2,7-dioxo-
2,3-
dihydro-1H,7H-pyrido[1,2,3-de]quinoxaline-6-carboxamide (0.21 g) and I-(2-
furyl)-
2-(methylamino)ethanol (0.10 g) is added DMF (5 mL) and diisopropylethylamine
(0.5 mL). After 3 days of stirring at room temperature, the reaction mixture
is diluted
with ethyl acetate (100 mL), washed with dilute pH 4 phosphate buffer (4 x 10
mL),
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brine, dried over sodium sulfate, filtered, and finally concentrated under
reduced
pressure. The residue is flash column chromatographed on silica eluting with
4% to
9% methanol in ethyl acetate. The product containing fractions are combined
and
concentrated under reduced pressure. The resulting residue is crystallized
from
toluene to yield 0.21 g of the title compound as a white solid. Physical
characteristics:
Mp 153-155 °C;1H NMR (d6-DMSO) 8 10.4, 8.73, 7.80, 7.55, 7.4, 7.3,
6.38, 6.28,
5.28, 5.22, 4.77, 4.57, 3.7, 3.34, 2.7, 2.21; HRMS (ESI+) mlz 535.1768 (M+H)~;
Anal. Found: C, 62.62; H, 5.05; N, 10.40.
l0 EXAMPLE 6 Preparation of rac N-(4-chlorobenzyl)-9-{([2-hydroxy-2-(5-methyl-
2-
furyl)ethyl](methyl)amino)methyl}- 1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-
pyrido[ 1,2,3-de]quinoxaline-6-carboxamide.
0 0
~N ~ I I NH I
OH ~ ~ N ~~I
~N~
(~O
To a flask containing N-(4-chlorobenzyl)-9-(chloromethyl)-1-methyl-2,7-dioxo-
2,3-
dihydro-1H,7H-pyrido[1,2,3-de]quinoxaline-6-carboxamide (0.14 g) and 2-
(methylamino)-1-(5-methyl-2-furyl)ethanol (0.08 g) is added DMF (3 mL) and
diisopropylethylamine (0.5 mL). After 4 days of stirring at room temperature,
the
2o reaction mixture is diluted with ethyl acetate ( 100 mL), washed with
dilute pH 4
phosphate buffer (4 x 10 mL), brine, dried over sodium sulfate, filtered, and
finally
concentrated under reduced pressure. The residue is adsorbed onto silica and
flash
column chromatographed on silica eluting with 3% to 9% methanol in ethyl
acetate.
The product containing fractions are combined and concentrated under reduced
pressure. The resulting residue is crystallized from toluene to yield 0.14 g
of the title
compound as a white solid. Physical characteristics: Mp 159-161 °C;1H
NMR (d6-
DMSO) ~ 10.4, 8.73, 7.80, 7.4, 6.12, 5.96, 5.22, 5.15, 4.7, 4.57, 3.7, 3.35,
2.70, 2.22,
2.18; HRMS (ESI+) m/z 549.1920 (M+H)+; Anal. Found: C, 63.32; H, 5.44; N,
10.08.
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EXAMPLE 7 Preparation of rac N-(4-chlorobenzyl)-9-{([2-(2,5-dimethyl-3-furyl)-
2-
hydroxyethylj(methyl)amino)methyl}- 1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-
pyrido [ 1,2,3-de] quinoxaline-6-carboxamide.
cl
To a flask containing N-(4-chlorobenzyl)-9-(chloromethyl)-1-methyl-2,7-dioxo-
2,3-
dihydro-1H,7H-pyrido[1,2,3-de]quinoxaline-6-carboxamide (0.14 g) and 1-(2,5-
dimethyl-3-furyl)-2-(methylamino)ethanol (0.08 g) is added DMF (3 mL) and
l0 diisopropylethylamine (0.5 mL). After 4 days of stirring at room
temperature, the
reaction mixture is diluted with ethyl acetate ( 100 mL), washed with dilute
pH 4
phosphate buffer (4 x 10 mL), brine, dried over sodium sulfate, filtered, and
finally
concentrated under reduced pressure. The residue is adsorbed onto silica and
flash
column chromatographed on silica eluting with 3% to 9% methanol in ethyl
acetate.
15 The product containing fractions are combined and concentrated under
reduced
pressure. The resulting residue is crystallized from toluene to yield 0.13 g
of the title
compound as a white solid. Physical characteristics: Mp 157-160 °C;1H
NMR (d6-
DMSO) 8 10.4, 8.73, 7.80, 7.4, 5.85, 5.22, 4.75, 4.57, 3.7, 3.35, 2.7-2.4,
2.22, 2.16,
2.13; HRMS (ESI+) yr~lz 563.2039 (M+H)+.
EXAMPLE 8 Preparation of rac N-(4-chlorobenzyl)-9-{([2-hydroxy-2-(6-
methylpyridin-2-yl)ethyl] (rnethyl)amino)methyl }-1-methyl-2,7-dioxo-2,3-
dihydro-
1 H,7H-pyrido [ 1,2, 3-dej quinoxaline-6-carboxamide.
i~ 0 0
N N i NH
OH
N CI
~N~
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To a flask containing N-(4-chlorobenzyl)-9-(chloromethyl)-1-methyl-2,7-dioxo-
2,3-
dihydro-1H,7H-pyrido[1,2,3-de]quinoxaline-6-carboxamide (0.14 g) is added DMF
(3
mL) followed by 2-(methylamino)-1-(6-methylpyridin-2-yl)ethanol (0.09 g) and
diisopropylethylamine (0.5 mL). After 4 days of stirring at room temperature,
the
reaction mixture is diluted with ethyl acetate (100 mL), washed with dilute pH
4
phosphate buffer (3 x 10 mL), brine, dried over sodium sulfate, filtered, and
finally
concentrated under reduced pressure. The residue is flash column
chromatographed on
silica eluting with 2% to 6% methanol in dichloromethane. The product
containing
fractions are combined and concentrated under reduced pressure. The resulting
residue
1o is crystallized from toluene to yield 0.15 g of the title compound as a
white solid.
Physical characteristics: Mp 156-158 °C;1H NMR (d6-DMSO) 8 10.4,
8.73, 7.79,
7.63, 7.4, 7.3, 7.1, 5.3, 5.22, 4.8, 4.56, 3.7, 3.35, 2.8-2.6, 2.39, 2.27;
HRMS (ESI+)
m/z 560.2041 (M+H)+; Anal. Found: C, 64.08; H, 5.52; N, 12.40.
EXAMPLE 9 Preparation of N-(4-chlorobenzyl)-9-{ [[(2R)-2-hydroxy-2-pyridin-2-
ylethyl] (methyl)amino]methyl }-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[
1,2,3-
de]quinoxaline-6 carboxamide.
~N~ N
OH H I ~ CI
A 25 mL 1-neck 14120 round bottom flask is charged with N-(4-chlorobenzyl)-9-
(chloromethyl)-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[I,2,3-
de]quinoxaline-
6-carboxamide (0.25 g) and R-2-(1-hydroxy-2-N-methylamino-ethyl)-pyridine
(0.177
g). To this mixture is added DMF (5mL), (i-Pr)~,NEt (0.15 g), and dried, 4~1
powedered molecular sieves ( 1.16 g). The mixture is allowed to stir at room
temperature for 16 hours. The mixture is diluted with CHZC12 (50 mL), filtered
through a pad of Celite~ on a coarse scintered glass funnel and the filter
cake is rinsed
with CHZCl2 (50 mL). The bulk of the solvent is removed on a rotary evaporator
and
the redisual DMF is removed at high vacuum to a yellow oil. Purification on a
40S
Biotage column [wet CH2Cl2, eluted CHZC12, CH2C12/MeOH (97.5:2.5),
CH2Cla/MeOH (95:5), CH2C12/MeOH (92.5:7.5), CHZC12/MeOH (90:10) provides
0.301 g of N-(4-chlorobenzyl)-9-{ [[(2R)-2-hydroxy-2-pyridin-3-
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ylethyl] (methyl)amino]methyl }-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[
1,2,3-
de]quinoxaline-6-carboxamide as an ivory solid. Physical characteristics: mp:
125-
127°C; 1H-NMR (400MHz, CDC13): 8 10.28, 8.68, 8.53, 8.00, 7.70, 7.46,
7.40, 7.31,
7.21, 5.02, 4.94, 4.64, 3.93, 3.73, 3.55, 2.88, 2.74, 2.42; 13C-NMR (100MHz,
CDCl3):
8 182.21, 173.84, 169.88, 162.18, 158.24, 146.58, 142.92, 135.07, 134.71,
130.84,
128.75, 126.89, 126.64, 125.16, 124.70, 118.72, 118.42, 115.56, 111.82, 68.23,
61.70,
59.97, 50.39, 40.59, 40.31, 27.12.; Specific Rotation [a]D2s 51 (c 0.41,
chloroform).
EXAMPLE 10 Preparation of rac N-(4-chlorobenzyl)-9-{([2-hydroxy-2-(4-hydroxy-
l0 3-methoxyphenyl)ethyl](methyl)amino)methyl}-1-methyl-2,7-dioxo-2,3-dihydro-
1 H,7H-pyrido [ 1, 2, 3-de] quinoxaline-6-carboxamide.
Ho ~ I o 0
~O~N i NH
OH
N CI
~N~
O
To a flask containing N-(4-chlorobenzyl)-9-(chlorornethyl)-1-methyl-2,7-dioxo-
2,3-
dihydro-1H,7H-pyrido[1,2,3-de]quinoxaline-6-carboxamide (0.14 g) and
metanephrine hydrochloride (0.12 g) is added DMF (3 mL) and
diisopropylethylamine
(1.0 mL). After 7 days of stirring at room temperature, the reaction mixture
is diluted
with ethyl acetate (100 rnL) and washed with dilute pH 7 phosphate buffer
(3x). The
combined aqueous layers are back extracted with ethyl acetate. The combined
organic
layers are washed with brine, dried over sodium sulfate, filtered, and finally
concentrated under reduced pressure. The residue is precipitated from methanol
-
toluene to yield 0.12 g of the title compound as an off white solid. Physical
characteristics: Mp 133-136 °C;1H NMR (d6-DMSO) 8 10.4, 8.74, 7.8, 7.4,
7.3, 6.8,
6.7, 5.21, 5.0, 4.7, 4.6, 3.6, 3.3, 2.6, 2.26; HRMS (ESI+) m/.z 591.2006
(M+H)+; Anal.
Found: C, 62.73; H, 5.36; N, 9.32.
Example 11 Preparation of N-(4-chlorobenzyl)-9-{ [[(2R)-2-(2-furyl)-2-
hydroxyethyl](methyl)amino]methyl }-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-
pyrido[I,2,3-de]quinoxaline-6-carboxamide.
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0 0
v TcH l I ~ I H I
ci
0
As described in the general procedure outlined in Example 9, a stirxing
solution of N-
methyl R-1-(2-furyl)-2-aminoethanol (0.16 g), powdered 4~ sieves (1.I6 g),
iPr2NEt
(0.2 mL) and N-(4-chlorobenzyl)-9-(chloromethyl)-1-methyl-2,7-dioxo-2,3-
dihydro-
1H,7H-pyrido[1,2,3-de]quinoxaline-6-carboxamide (0.25 g) in DMF (5 mL) affords
a
crude a yellow oil. The crude material is chromatographed on a 40g Biotage
column
(packed with CHZC12; eluted with CH2C12, then 5% MeOH/CH2C12 to afford N-(4-
l0 chlorobenzyl)-9-{[[(2R)-2-(2-furyl)-2-hydroxyethyl](methyl)amino] methyl}-1-
methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[1,2,3-de]quinoxaline-6-cwboxamide as
a white solid (0.16 g). Physical characteristics: Mp119-120°C;1H-NMR
(300MHz,
CDC13): 8 10.28, 8.65, 8.00, 7.22-7.50, 6.34, 6.29, 5.00, 4.85, 4.65, 3.88,
3.72, 3.53,
3.48, 3.00, 2.70, 2.40; HRMS (FAB) found 535.1733 for C2sH27CLN405+H+;
Specific
15 Rotation [oc]25D = 23 (c 0.98, chloroform).
Example 12 Preparation of N-(4-chlorobenzyl)-9-{ [[(2S)-2-hydroxy-2-
phenylethyl] (methyl)amino]methyl }-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-
pyrido[ 1,2,3-de]quinoxaline-6-carboxamide.
I o 0
N I ~ I H I ~
OH I i N ~CI
,N\ J
~O
a
As described in the general procedure outlined in Example 9, powdered 4A
sieves
(1.8 g), iPr2NEt (0.2 mL) and N-(4-chlorobenzyl)-9-(chloromethyl)-1-methyl-2,7-
dioxo-2,3-dihydro-1H,7H-pyrido[1,2,3-de]quinoxaline-6-carboxamide (0.40 g) and
S-
2-(1-hydroxy-2-N-methylamino-ethyl)-benzene (0.25 g) in DMF (10 mL) affords
0.30
g of N-(4-chlorobenzyl)-9-{[[(2S)-2-hydroxy-2-
phenylethyl](methyl)amino]methyl}-
1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[1,2,3-de]quinoxaline-6-carboxamide
as
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a pale yellow oil. Physical characteristics: 1H-NMR (300MHz, DMSO-d6): b
10.35,
8.73, 7.95, 7.13-7.42, 5.21, 5.16, 4.76, 4.56, 3.69, 3.30-3.40, 2.63, 2.49,
2.25; HRMS
(FAB) found 545.1963 for C3nH29CLN4O4 +Hl; Specific Rotation [a]25D = 11 (c
1.00,
DMSO).
Example 13 Preparation of N-(4-chlorobenzyl)-9-{ [[(2S)-2-hydroxy-2-pyridin-3-
ylethyl] (methyl)amino]methyl }-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[
1,2,3-
de] quinoxaline-6-carboxamide.
0 0
N~ I N
OH I I ~ I H I ~
N CI
,N\ J
As described in the general procedure outlined in Example 9, 0.40 g of N-(4-
chlorobenzyl)-9-(chloromethyl)-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-
pyrido[1,2,3-
de]quinoxaline-6-carboxamide, 0.28 g of S-3-(1-hydroxy-2-N-methylamino-ethyl)-
pyridine, 0.30 mL of i-PrzNEt, and 1.8 g of 4A sieves in DMF (10 mL,) affords
0.438
of N-(4-chlorobenzyl)-9-{ [[(2S)-2-hydroxy-2-pyridin-3- .
ylethyl] (methyl)amino]methyl }-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[
1,2,3-
de]quinoxaline-6-carboxamide as an ivory solid. Physical characteristics: mp:
131-
135°C; rH-NMR (300MHz, DMSO-d6): 8 10.35, 8.72, 8.52, 8.41, 7.75, 7.68,
7.25-
7.47, 7.23, 5.37, 5.29, 4.81, 4.56, 3.71, 3.62, 3.29, 2.67, 2.57, 2.25; HRMS
(FAB)
found 546.1910 for C29H28CLN504 +Hl ; Specific Rotation [a]25D = 7 (c 1.03,
DMSO).
Example 14 N-(4-chlorobenzyl)-9-{ [[(2S)-2-hydroxy-2-thien-3-
ylethyl](methyl)amino]methyl}-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-
pyrido[1,2,3-
de]quinoxaline-6-carboxamide.
0 0
OH I I ~ I H I
N CI
N\ J
~O
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As described in the general procedure outlined in Example 9, 0.40 g of N-(4-
chlorobenzyl)-9-(chloromethyl)-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-
pyrido[1,2,3-
de]quinoxaline-6-carboxamide, 0.28 g of S-3-(1-hydroxy-2-N-methylamino-ethyl)-
thiophene, 0.30 mL of i-Pr2NEt, and 1.8 g of 4A sieves in DMF (10 mL) affords
0.48
g of N-(4-chlorobenzyl)-9-{ [[(2S)-2-hydroxy-2-thien-3-ylethyl](methyl)
amino] methyl } -1-methyl-2,7-dioxo-2, 3-dihydro-1 H,7H-pyrido [ 1,2,3-de]
quinoxaline-
6-carboxamide as an ivory solid. Physical characteristics: 1H-NMR (300MHz,
DMSO-d6): 8 10.35, 8.72, 7.79, 7.25-7.46, 7.06, 5.21, 5.15, 4.85, 4.56, 3.69,
3.37,
3.34, 2.64, 2.24; mp 137-139°C; HRMS (FAB) found 551.1530 for
C28H2~CLN~O4S
l0 +Hl ; Specific Rotation [a]25D = 40 (c 1.01, chloroform).
EXAMPLE 15 Preparation of rac 9-{([2-(1-benzothien-3-yl)-2-
hydroxyethyl](methyl)amino)methyl}-N-(4-chlorobenzyl)-1-methyl-2,7-dioxo-2,3-
dihydro-1H,7H-pyrido[ 1,2,3-de]quinoxaline-6-carboxamide.
To a flask containing N-(4-chlorobenzyl)-9-(chloromethyl)-1-methyl-2,7-dioxo-
2,3-
dihydro-1H,7H-pyrido[1,2,3-de]quinoxaline-6-carboxamide (0.14 g) and 1-(1-
benzothien-3-yl)-2-(methylamino)ethanol (0.10 g) is added DMF (3 mL) and
diisopropylethylamine (0.5 mL). After 4 days of sthTing at room temperature,
the
reaction mixture is diluted with ethyl acetate (100 mL), washed with dilute pH
4
phosphate buffer (4 x 10 mL), brine, dried over sodium sulfate, filtered, and
finally
concentrated under reduced pressure. The residue is adsorbed onto silica and
flash
column chromatographed on silica eluting with 3% to 9% methanol in ethyl
acetate.
The product containing fractions are combined and concentrated under reduced
pressure. The resulting residue is crystallized from toluene and then
recrystallized
from acetonitrile - methanol to yield 0.12 g of the title compound as a white
solid.
Physical characteristics: Mp 147-149 °C;1H NMR (d6-DMSO) 8 10.4,
7.98, 7.77,
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7.68, 7.54, 7.4, 7.3-7.1, 5.35, 5.21, 5.1, 4.9, 4.57, 3.7, 3.22, 2.9-2.6,
2.37; HRMS
(ESI+) fnlz 601.1657(M+H)+; Anal. Found: C, 63.83; H, 4.93; N, 9.30.
EXAMPLE 16 Preparation of rac N-(4-chlorobenzyl)-9-{([2-hydroxy-2-quinolin-2-
ylethyl](methyl)amino)methyl}-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-
pyrido[1,2,3-
de]quinoxaline-6-carboxamide.
0 0
N N ~ NH ~
OH ~ W ( N I ~CI
~N~
nO
To a flask containing N-(4-chlorobenzyl)-9-(chloromethyl)-1-methyl-2,7-dioxo-
2,3-
dihydro-1H,7H-pyrido[1,2,3-de]quinoxaline-6-carboxamide (0.14 g) is added DMF
(3
mL) followed by 2-(methylamino)-1-quinolin-2-ylethanol (0.10 g) and
diisopropylethylamine (0.5 mL). After 6 days of stirring at room temperature,
the
reaction mixture is diluted with ethyl acetate (90 mL) and washed with dilute
pH 4
phosphate buffer (3 x 10 mL). The combined aqueous layers are back extracted
with
ethyl acetate and dichloromethane. The combined organic layers are washed with
brine, dried over sodium sulfate, filtered, and finally concentrated under
reduced
pressure. The residue is flash column chromatographed on silica eluting with
3% to
7.5% methanol in dichloromethane. The product containing fractions are
combined
and concentrated under reduced pressure. The resulting residue is crystallized
from
toluene to yield 0.11 g of the title compound as a white solid. Physical
characteristics:
Mp 173-177 °C;1H NMR (d6-DMSO) 8 10.4, 8.71, 8.28, 7.91, 7.85, 7.71,
7.66, 7.58,
7.55, 7.4, 7.04, 5.55, 5.16, 4.9, 4.57, 3.7, 3.01, 2.9-2.7, 2.31; HRMS (ESI+)
m/z
596.2068 (M+H)+; Anal. Found: C, 66.41; H, 5.17; N, 11.74.
EXAMPLE 17 Preparation of rac N-(4-chlorobenzyl)-9-{ [(2-hydroxy-3-
phenylpropyl)(methyl)amino]methyl }-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-
pyrido [ 1, 2, 3-de] quinoxaline-6-carboxamide.
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I
O O
OH ~ ~ I ( NH
~N' CI
~N~
nO
To a flask containing N-(4-chlorobenzyl)-9-(chloromethyl)-1-methyl-2,7-dioxo-
2,3-
dihydro-1H,7H-pyrido[1,2,3-de]quinoxaline-6-carboxamide (0.14 g) and 1-
(methylamino)-3-phenylpropan-2-of (0.08 g) is added DMF (3 mL) and
diisopropylethylamine (0.5 mL). After 7 days of stirring at room temperature,
the
reaction mixture is diluted with ethyl acetate (60 mL) and washed with dilute
pH 4
phosphate buffer (3x). The combined aqueous layers are back extracted with
ethyl
acetate. The combined organic layers are washed with brine, dried over sodium
sulfate, filtered, and finally concentrated under reduced pressure. The
residue is
crystallized from acetonitrile to yield 0.14 g of the title compound as an off
white
solid. Physical characteristics: mp 121-127 °C;1H NMR (d~-DMSO) 810.4,
8.74,
7.85, 7.46, 7.4, 7.3-7.1, 5.23, 4.6, 3.9, 3.66, 3.3, 2.8, 2.5, 2.34, 2.22;
HRMS (ESI+)
fnlz 559.2131 (M+H)~; Anal. Found: C, 66.44; H, 5.80; N, 9.83.
EXAMPLE 18 Preparatio of rac N-(4-chlorobenzyl)-9-{([2-hydroxy-2-(3-
methoxyphenyl)ethyl] (methyl)amino)methyl }-1-methyl-2,7-dioxo-2,3-dihydro-
I H,7H-pyrido [ I ,2,3-de] quinoxaline-6-carboxarnide.
iI o 0
~O~N ~ NH
OH ~ ~ I I
N CI
~N~
nO
To a flask containing N-(4-chlorobenzyl)-9-(chloromethyl)-1-methyl-2,7-dioxo-
2,3-
dihydro-1H,7H-pyrido[1,2,3-de]quinoxaline-6-carboxamide (0.14 g) and 1-(3-
methoxyphenyl)-2-(methylamino)ethanol (0.09 g) is added DMF (3 mL) and
diisopropylethylamine (0.5 mL). After 7 days of stirring at room temperature,
the
reaction mixture is diluted with ethyl acetate (60 mL), washed with dilute pH
4
phosphate buffer (4x), brine, dried over sodium sulfate, filtered, and finally
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concentrated under reduced pressure. The residue is crystallized from
acetonitrile. The
crystals are adsorbed onto silica and flash column chromatographed on silica
eluting
with 2% to 6% methanol in dichloromethane. The product containing fractions
are
combined and concentrated under reduced pressure. The resulting residue is
crystallized from acetonitrile to yield 0.06 g of the title compound as a
white solid.
Physical characteristics: Mp 154-156 °C;1H NMR (d6-DMSO) b 10.4, 8.74,
7.80, 7.4,
7.2, 6.9, 6.8, 5.22, 5.15, 4.75, 4.57, 3.7, 3.3, 2.6, 2.26; HRMS (ESI+) m/z
575.2068
(M+H)+; Anal. Found: C, 64.49; H, 5.42; N, 9.69.
to EXAMPLE 19 Preparation of rac N-(4-chlorobenzyl)-9-{([2-hydroxy-2-pyrimidin-
2-
ylethyl] (methyl)amino)methyl }-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido [
1,2,3-
de] quinoxaline-6-carboxamide.
~N O O
JI~
N off ~ ~ I I NH ~ I
'N' CI
~N~
nO
To a flask containing N-(4-chlorobenzyl)-9-(chloromethyl)-1-methyl-2,7-dioxo-
2,3-
dihydro-1H,7H-pyrido[1,2,3-de]quinoxaline-6-carboxamide (0.27 g) and 2-
(methylamino)-1-pyrimidin-2-ylethanol (0.15 g) is added DMF (5 mL) and
diisopropylethylamine (1.0 mL). After 4 days of stirring at room temperature,
the
reaction mixture is diluted with ethyl acetate (100 mL), washed with dilute pH
4
phosphate buffer (2 x 50 mL), brine, dried over sodium sulfate, filtered, and
finally
concentrated under reduced pressure. The residue is flash column
chromatographed on
silica eluting with 2% to 6% methanol in dichloromethane. The product
containing
fractions are combined and concentrated under reduced pressure. The resulting
residue
is crystallized from acetonitrile to yield 0.27 g of the title compound as a
white solid.
Physical characteristics: Mp 168-170 °C;1H NMR (d6-DMSO) ~ 10.4,
8.77, 8.75,
8.72, 7.71, 7.4, 7.20, 5.32, 5.21, 4.85, 4.57, 3.7, 3.3, 2.9, 2.7, 2.21; HRMS
(ESI+) m/z
547.1853 (M+H)~; Anal. Found: C, 61.16; H, 4.96; N, 15.30.
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EXAMPLE 20 Preparation of rac N-(4-chlorobenzyl)-9-{([2-(5-cyanothein-2-yl)-2-
hydroxyethyl](methyl)amino)methyl}- 1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-
pyrido[ 1,2,3-de]quinoxaline-6-carboxamide.
0 0
S OH ~ ~ I I NH ~ I
~N~ CI
O
To a flask containing N-(4-chlorobenzyl)-9-(chloromethyl)-1-methyl-2,7-dioxo-
2,3-
dihydro-1H,7H-pyrido[1,2,3-de]quinoxaline-6-carboxamide (0.27 g) and 5-[1-
hydroxy-2-(methylamino)ethyl]thiophene-2-carbonitrile (0.18 g) is added DMF (5
mL) and diisopropylethylamine (1.0 mL). After 4 days of stirring at room
temperature,
the reaction mixture is diluted with ethyl acetate ( 100 mL), washed with
dilute pH 4
phosphate buffer (2 x 50 mL), brine, dried over sodium sulfate, filtered, and
finally
concentrated under reduced pressure. The residue is flash column
chromatographed on
silica eluting with 2% to 4% methanol in dichloromethane. The product
containing
fractions are combined and concentrated under reduced pressure. The resulting
residue
is crystallized from acetonitrile - toluene -methanol to yield 0.23 g of the
title
compound as a white solid. Physical characteristics: Mp 122-126 °C;1H
NMR (d6-
DMSO) 8 10.4, 8.73, 7.81, 7.4, 7.14, 6.08, 5.21, 5.06, 4.56, 3.7, 3.35, 2.7,
2.26;
HRMS (ESI+) rnlz 576.1486 (M+H)+; % Water (KF): 0.53; Anal. Found: C, 59.90;
H,
4.65; N, 12.07.
Example 21 Preparation of N-(4-chlorobenzyl)-9-{ [[(2R)-2-hydroxy-2-pyrazin-2
ylethyl] (methyl)amino]methyl }-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido [
1,2,3
de]quinoxaline-6-carboxamide.
O O
~I ~
CN~ I I \ I H I \
OH / N ~CI
,N\ J
~(O
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As described in the general procedure outlined in Example 9, 0.32 g of N-(4-
chlorobenzyl)-9-(chloromethyl)-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-
pyrido[1,2,3-
de]quinoxaline-6-carboxamide, 0.23 g of R-2-(1-hydroxy-2-N-methylamino-ethyl)-
pyrazine, 0.25 mL of i-Pr2NEt, and 0.5 g of 4A sieves in DMF (5 mL) affords
0.3 I g
of N-(4-chlorobenzyl)-9-{[[(2R)-2-hydroxy-2-pyrazin-2-ylethyl](methyl)
amino] methyl } -1-methyl-2,7-dioxo-2,3-dihydro-1 H,7H-pyrido [ 1,2, 3-de]
quinoxaline-
6-carboxamide as an ivory solid. Physical characteristics: mp 97-
98°C;1H-NMR
(400MHz, DMSO-d6): 8 I0.37, 8.71, 8.51, 7.72, 7.32-7.43, 7.21, 5.62, 5.21,
4.88,
4.56, 3.67, 3.30, 2.83, 2.74, 2.24; HRMS (FAB) found 547.1880 fox C28H27CLN6O4
io +Hl ; Specific Rotation [a]ZSD = 23 (c 0.9I, DMSO).
Example 22 Preparation of N-(4-chlorobenzyl)-9-{ [[(2R)-2-hydroxy-2-(1,3-
thiazol-
2-yl)ethyl] (methyl)amino]methyl }-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-
15 pyrido[1,2,3-de]quinoxaline-6-carboxamide.
~s o 0
N N I ~ I H I ~
I N~
,N\ J
~O
As described in the general procedure outlined in Example 9, 0.46 g of N-(4-
2o chlorobenzyl)-9-(chloromethyl)-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-
pyrido[I,2,3-
de]quinoxaline-6-carboxamide, 0.34 g of R-2-(1-hydroxy-2-N-methylamino-ethyl)-
0
thiazole, 0.37 mL of i-Pr2NEt, and 0.5 g of 4A sieves in DMF (5 mL) affords
0.27 g
of N-(4-chlorobenzyl)-9-{ [[(2R)-2-hydroxy-2-(1,3-thiazol-2-
yl)ethyl](methyl)amino]
methyl}-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[1,2,3-de]quinoxaline-6-
25 carboxamide as an ivory solid. Physical characteristics: mp125-
130°C;1H-NMR
(400MHz, DMSO-d6): 8 10.36, 8.72, 7.80, 7.72, 7.62, 7-30-7.42, 6.I8, 5.76,
5.2I,
5.07, 4.56, 3.73, 3.36, 2.85, 2.74, 2.26; HRMS (FAB) found 552.1469 for
C27H26CLN504S +Hl ; Specific Rotation [a]25D = 12 (c 0.96, DMSO).
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Example 23 Preparation of N-(4-fluorobenzyl)-9-{ [[(2R)-2-(2-furyl)-2-
hydroxyethyl](methyl)amino]methyl}-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-
pyrido[1,2,3-de]quinoxaline-6-carboxamide.
0 0
0 OH I I ~ I H I
N F
~N
O
As described in the general procedure outlined in Example 9, 0.25 g of N-(4-
fluorobenzyl)-9-(chloromethyl)-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-
pyrido[1,2,3-
de]quinoxaline-6-carboxamide, 0.17 g of N-Methyl R-1-(2-furyl)-2-aminoethanol,
0.21 mL of i-Pr2NEt, and 0.3 g of 4A sieves in DMF (5 mL) affords 0.18 g of N-
(4-
fluorobenzyl)-9-{ [[(2R)-2-(2-furyl)-2-hydroxyethyl](methyl)amino]methyl}-1-
methyl-
2,7-dioxo-2,3-dihydro-1H,7H-pyrido[1,2,3-de]quinoxaline-6-carboxarnide as an
ivory
solid. Physical characteristics: mp 150-152°C; 1H-NMR (400MHz, DMSO-
d6):
S 10.33, 8.72, 7.78, 7.44, 7.39, 7.26, 7.16, 6.33, 6.27, 5.27, 5.21, 4.75,
4.55, 3.66,
3.37, 2.71, 2.23; HRMS (FAB) found 519.2043 for C28H27FN4O5 +HI ; Specific
Rotation [a]ZSD = -2 (c 0.97, DMSO).
Example 24 Preparation of N-(4-fluorobenzyl)-9-{ [[(2R)-2-hydroxy-2-pyridin-2-
ylethyl] (methyl)amino]methyl }-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido [
1,2,3-
de]quinoxaline-6-carboxamide.
( ~ 0 0
N N I ~ I H
OH ~ ~ N ~F
~N\J
[JO
As described in the general procedure outlined in Example 9, 0.25 g of N-(4-
fluorobenzyl)-9-(chloromethyl)-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-
pyrido[1,2,3-
de]quinoxaline-6-carboxamide, 0.18 g of N-methyl R-2-(1-hydroxy-2-N-
methylamino-ethyl)-pyridine, 0.21 mL of i-PrzNEt, and 0.30 g of 41~ sieves in
DMF
(5 mL) affords 0.16 g of N-(4-fluorobenzyl)-9-{ [[(2R)-2-hydroxy-2-pyridin-2-
ylethyl] (methyl)amino]methyl }-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[
1,2,3
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de]quinoxaline-6-carboxamide as a pale yellow solid. Physical characteristics:
mp
121-123; 1H-NMR (400MHz, DMSO-db): 8 10.35, 8.73, 8.45, 7.70-7.80, 7.46, 7.30-
7.40, 7.22, 7.16, 5.36, 5.21, 4.82, 4.55, 3.71, 3.33, 2.75, 2.66, 2.26; HRMS
(FAB)
found 530.2184 for C29H28FNSO4 +Hl ; Specific Rotation [a]25D = 27 (c 0.60,
DMSO).
Example 25 Preparation of N-(4-fluorobenzyl)-9-{ [[(2S)-2-hydroxy-2-pyridin-3-
ylethyl] (methyl)amino]methyl }-1-methyl-2,7-dioxo-2,3-dihydro-1 H,7H-pyrido [
1,2,3-
de] quinoxaline-6-carboxamide.
to
0 0
Nr
OH ~ I ~ I H
N F
~N\J
~O
As described in the general procedure outlined in Example 9, 0.25 g of N-(4-
fluorobenzyl)-9-(chloromethyl)-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[
1,2;3-
de]quinoxaline-6-carboxamide, 0.18 g of S-3-(1-hydroxy-2-N-methylamino-ethyl)-
pyridine, 0.21 mL of i-PrZNEt, and 0.30 g of 4~ sieves in DMF (5 rnL) affords
0.23 g
of N-(4-fluorobenzyl)-9-{ [[(2S)-2-hydroxy-2-pyridin-3-ylethyl](methyl)
amino]methyl }-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[ 1,2,3-
de]quinoxaline-
6-carboxamide as a pale yellow solid. Physical characteristics: mp 108-
110°C; 1H-
2o NMR (400MHz, DMSO-d6): 8 10.21, 8.75, 8.51, 8.41, 7.75, 7.68, 7.32, 7.31,
7.22,
7.16, 5.37, 5.21, 4.81, 4.55, 3.70, 3.62, 3.34, 2.66, 2.57, 2.24; HRMS (FAB)
found
530.2196 for C29H28 '~5~4 '~'Hl ; Specific Rotation [a]25D = 2 (c 0.90, DMSO).
EXAMPLE 26 Preparation of rac 9-{([2-(1-benzofuran-2-yl)-2-
hydroxyethyl](methyl)amino)methyl}-N-(4-chlorobenzyl)-1-(2-hydroxyethyl)-2,7-
dioxo-2,3-dihydro-1 H,7H-pyrido [ 1,2, 3-de] quinoxaline-6-carboxamide.
0 0
"~N ~ I NH ~
OH ~ ~ N ~ v'CI
HON
O
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To a flask containing N-(4-chlorobenzyl)-9-(chloromethyl)-1-(2-hydroxyethyl)-
2,7-
dioxo-2,3-dihydro-1H,7H-pyrido[1,2,3-de]quinoxaline-6-carboxamide (0.20 g) and
1-(1-benzofuran-2-yl)-2-(methylamino)ethanol (0.13 g) is added DMF (3.5 mL)
and
diisopropylethylamine (0.7 mL). After stirring at room temperature overnight,
the
reaction mixture is diluted with ethyl acetate (75 mL), washed with dilute pH
4
phosphate buffer (25 mL), dilute pH 7 phosphate buffer (25 mL), brine, dried
over
sodium sulfate, filtered, and finally concentrated under reduced pressure. The
residue
is adsorbed onto silica and flash column chromatographed on silica eluting
with 3% to~
8% methanol in dichloromethane. The product containing fractions are combined
and
concentrated under reduced pressure. The resulting residue is crystallized
from
methanol - acetonitrile to yield 0.15 g of the title compound as a white
solid. Physical
characteristics: Mp 195-197 °C;1H NMR (d6-DMSO) 8 10.4, 8.73, 7.79,
7.55, 7.4,
7.2, 6.73, 5.53, 5.18, 4.85, 4.57, 3.93, 3.8-3.6, 3.48, 2.9, 2.7, 2.29; HRMS
(ESI+) m/z
615.2008 (M+H)+.
Example 27 Preparation of N-(4-fluorobenzyl)-9-{ [[(2R)-2-hydroxy-2-(1,3-
thiazol-
2-yl)ethyl] (methyl)amino]methyl }-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-
pyrido[ 1,2,3-de] quinoxaline-6-carboxamide.
0 0
OH N I / I H I
I NJ F
~N~
As described in the general procedure outlined in Example 9, 0.25 g of N-(4-
fluorobenzyl)-9-(chloromethyl)-1-methyl-2,7-dioxo-2,3-dihydro-1 H,7H-pyrido [
1,2, 3-
de]quinoxaline-6-carboxamide, 0.19 g of R-2-(1-hydroxy-2-N-methylamino-ethyl)-
thiazole, 0.21mL of i-Pr2NEt, and 0.30 g of 4~ sieves in DMF (5 mL) affords
0.21 g
of N-(4-fluorobenzyl)-9-{ [[(2R)-2-hydroxy-2-(1,3-thiazol-2-
yl)ethyl](methyl)amino]
methyl }-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[ 1,2,3-de]quinoxaline-6-
carboxamide as a pale yellow solid. Physical characteristics: mp 156-
158°C; 1H-NMR
(400MHz, DMSO-d6): 8 10.35, 8.73, 7.79, 7.22, 7.62, 7.35-7.45, 7.16, 6.18,
5.21,
5.06, 4.55, 3.61, 3.36, 2.85, 2.74, 2.26; HRMS (FAB) found 536.1750 for
C27H26~5~45 +Hl ; Specific Rotation [a]25D = 13 (c 0.99, DMSO).
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Example 28 Preparation of N-(4-fluorobenzyl)-9-{ [[(2S)-2-hydroxy-2-thien-3-
ylethyl] (methyl)axnino]methyl }-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[
1,2,3-
de] quinoxaline-6-carboxamide.
0 0
OH I I ~ I H I ~
N F
~N~
~O
As described in the general procedure outlined in Example 9, 0.25 g of N-(4-
fluorobenzyl)-9-(chloromethyl)-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[
1,2,3-
l0 de]quinoxaline-6-carboxamide, 0.19 g of S-3-(1-hydroxy-2-N-methylamino-
ethyl)-
thiophene, 0.21 mL of i-Pr2NEt, and 0.30 g of 4A sieves in DMF (5 mL) affords
0.20
g of N-(4-fluorobenzyl)-9-{ [[(2S)-2-hydroxy-2-thien-3-
ylethyl] (methyl)amino]methyl }-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[
1,2,3-
de]quinoxaline-6-carboxamide as a pale yellow solid. Physical characteristics:
mp
~5 15D-152°C; 1H-NMR (40DMHz, DMSO-d6): b 10.33, 8.73, 7.95, 7.80,
7.43, 7.35-
7.42, 7.31, 7.17, 7.05, 5.21, 5.14, 4.85, 4.55, 3.69, 3.35, 2.63, 2.24; HRMS
(FAB)
found 535.1799 for CZ8H27FN4O4S +Hl ; Specific Rotation [a.]25D = 6 (c 1.00,
DMSO).
20 Example 29 Preparation of N-(4-fluorobenzyl)-9-{ [[(2S)-2-hydroxy-2-
phenylethyl] (methyl)amino]methyl }-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-
pyrido [ 1,2,3-de]quinoxaline-6-carboxamide.
0 0
off I I i ~ "
~N' F
/N\ J
~O
As described in the general procedure outlined in Example 9, 0.25 g of N-(4-
fluorobenzyl)-9-(chloromethyl)-I-methyl-2,7-dioxo-2,3-dihydro-1H,7H-
pyrido[I,2,3-
de]quinoxaline-6-carboxamide, 0.18 g of S-a-[(methylamino)methyl]-
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benzenemethanol, 0.21 mL of i-Pr2NEt, and 0.30 g of 4A sieves in DMF (5 mL)
affords 0.20 g of N-(4-fluorobenzyl)-9-{ [[(2S)-2-hydroxy-2-
phenylethyl](methyl)amino] methyl}-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-
pyrido[1,2,3-de]quinoxaline-6-carboxamide as a pale yellow solid. Physical
characteristics: mp 150-152°; 1H-NMR (400MHz, DMSO-d6): 8,10.32, 8.73,
7.95,
7.86, 7.12-7.42, 5.21, 5.15, 4.76, 4.55, 3.69, 3.38, 3.31, 2.60, 2.53, 2.25;
HRMS
(FAB) found 529.2239 for C3pH29~4~4 "'f'Hl ; Specific Rotation [cc]25D = 10 (c
0.94,
DMSO).
l0 Example 30 Preparation of N-(4-fluorobenzyl)-9-{ [[(2R)-2-hydroxy-2-pyrazin-
2-
ylethyl] (methyl)amino]methyl }-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[
1,2,3-
de] quinoxaline-6-carboxamide.
CN I 0 O
\N~ N I \ ~ H
OH ~ N ~F
~ N\ J
~O
As described in the general procedure outlined in Example 9, 0.25 g of N-(4-
fluorobenzyl)-9-(chloromethyl)-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[
1,2,3-
de]quinoxaline-6-carboxamide, 0.18 g of R-2-(1-hydroxy-2-N-methylamino-ethyl)-
pyrazine, 0.21 mL of i-Pr2NEt, and 0.30 g of 4~ sieves in DMF (5 mL) affords
0.28 g
of N-(4-fluorobenzyl)-9-{ [[(2R)-2-hydroxy-2-pyrazin-2-
ylethyl] (methyl)amino]methyl }-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[
1,2,3-
de]quinoxaline-6-carboxamide as an ivory solid. Physical characteristics: mp
108-
110°C; 1H-NMR (400MHz, DMSO-d6): 8 10.35, 8.72, 8.70, 8.50, 7.25, 7.38,
7.21,
7.16, 5.62, 5.21, 4.87, 4.55, 3.67, 3.34, 3.31, 2.82, 2.73, 2.24; HRMS (FAB)
found
531.2152 for C28H27FN6O4 +Hl ; Specific Rotation [a,]ZSD = 25 (c 0.94, DMSO).
EXAMPLE 31 Preparation of N-(4-chlorobenzyl)-9-{ ([(2R)-2-(2-furyl)-2-
hydroxyethyl](methyl)amino)methyl}-1-(2-hydroxyethyl)-2,7-dioxo-2,3-dihydro-
1H,7H-pyrido[1,2,3-de]quinoxaline-6-carboxamide.
-44-
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WO 03/053971 PCT/US02/37613
/! O O
0 off ~ ~ i I NH
~N' CI
HON
O
To a flask containing N-(4-chlorobenzyl)-9-(chloromethyl)-1-(2-hydroxyethyl)-
2,7-
dioxo-2,3-dihydro-1H,7H-pyrido[1,2,3-de]quinoxaline-6-carboxamide (0.16 g) and
(1R)-1-(2-furyl)-2-(methylamino)ethanol (0.07 g) is added DMF (3 mL) and
diisopropylethylamine (0.6 mL). After stirring at room temperature for 12
days, the
reaction mixture is diluted with ethyl acetate (60 mL), washed with dilute pH
4
phosphate buffer (2 x 25 mL), brine, dried over sodium sulfate, filtered, and
finally
concentrated under reduced pressure. The residue is adsorbed onto silica and
flash
to column chromatographed on silica eluting with 3°lo to 9°~o
methanol in
dichloromethane. The product containing fractions are combined and
concentrated
under reduced pressure. The resulting residue is crystallized from
acetonitrile to yield
0.13 g of the title compound as a tan solid. Physical characteristics: Mp 166-
167 °C;
1H NMR (d6-DMSO) S 10.4, 8.74, 7.80, 7.55, 7.48, 6.38, 6.27, 5.24, 5.21, 4.88,
4.75,
4.56, 4.1, 3.7-3.6, 2.7, 2.21; HRMS (ESI+) m/z 565.1866 (M+H)+; Anal. Found:
C,
61.46; H, 5.24; N, 9.99.
Example 32 Preparation of N-(4-fluorobenzyl)-9-{[[(2R)-2-hydroxy-2-thien-2-
ylethyl] (methyl)amino]methyl }-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[
1,2,3-
de]quinoxaline-6-carboxamide.
0 0
S OH I I / I H I /
'N' F
/N\ J
~O
As described in the general procedure outlined in Example 9, 0.25 g of N-(4-
fluorobenzyl)-9-(chloromethyl)-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-
pyrido[1,2,3-
de]quinoxaline-6-carboxamide, 0.19 g of R-2-(1-hydroxy-2-N-methylamino-ethyl)-
thiophene, 0.21 mL of i-PrzNEt, and 0.30 g of 4A sieves in DMF (5 rnL) affords
0.19
g of N-(4-fluorobenzyl)-9-{ [[(2R)-2-hydroxy-2-thien-2-ylethyl](methyl)
- 45 -
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amino]methyl }-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[ 1,2,3-
de]quinoxaline-
6-carboxamide as an ivory solid. Physical characteristics: mp 113-
115°C; 1H-NMR
(400MHz, DMSO-d6): 8 10.32, 8.73, 7.95, 7.80, 7.30-7.40, 7.16, 6.99, 6.95,
5.56,
5.21, 4.96, 4.55, 3.75, 3.68, 3.34, 2.66, 2.60, 2.24; HRMS (FAB) found
535.1826 for
C28H27~4~4s '1'Hl.
Example 33 Preparation of rac 9-{ [[2-(1-benzofuran-2-yl)-2-
hydroxyethyl] (methyl)amino]methyl }-N-(4-fluorobenzyl)-1-methyl-2,7-dioxo-2,3-
dihydro-1 H,7H-pyrido [ 1,2,3-de] quinoxaline-6-carboxamide
\ / \ o 0
I ~ NI H
~N~
~O
As described in the general procedure outlined in Example 9, 0.25 g of N-(4-
fluorobenzyl)-9-(chloromethyl)-1-methyl-2,7-dioxo-2, 3-dihydro-1 H,7H-pyrido [
1,2, 3-
de]quinoxaline-6-carboxamide, 0.23 g of (~)-1-(1-benzofuran-2-yl)-2-
(methylamino)ethanol, 0.21 mL of i-Pr2NEt, and 0.30 g of 4~ sieves in DMF (5
mL)
affords 0.19 g of rac 9-{[[2-(1-benzofuran-2-yl)-2-hydroxyethyl](methyl)amino]
methyl }-N-(4-fluorobenzyl)-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[ 1,2,3-
de]quinoxaline-6-carboxamide as a white solid. Physical characteristics: mp161-
163°C; 1H-NMR (400MHz, DMSO-d6): 8 10.35, 8.72, 7.76, 7.53, 7.30-7.42,
7.10-
7.22, 6.73, 5.55, 5.17, 4.87, 4.55, 3.68, 3.60, 3.35, 2.88, 2.69, 2.29; HRMS
(FAB)
found 569.2214 for C3~Ha9FNøOS +Hl.
Example 34 Preparation of 9-{[[(2R)-2-(1-benzofuran-2-yl)-2-
hydroxyethyl](methyl)amino]methyl}-N-(4-chlorobenzyl)-1-methyl-2,7-dioxo-2,3-
dihydro-1 H,7H-pyrido [ 1,2,3-de]quinoxaline-6-carboxamide.
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O O
~~i' I ~ I H
N' CI
O
As described in the general procedure outlined in Example 9, 0.56 g of N-(4-
chlorobenzyl)-9-(chloromethyl)-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-
pyrido[1,2,3-
de]quinoxaline-6-carboxamide, 0.50 g of (R)-1-(1-benzofuran-2-yl)-2-
(methylamino)ethanol, 0.46 mL of i-Pr2NEt, and 0.65 g of 4~ sieves in DMF (35
mL)
affords 0.69g of 9-{[[(2R)-2-(1-benzofuran-2-yl)-2-hydroxyethyl](methyl)amino]
methyl }-N-(4-chlorobenzyl)-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[ 1,2,3-
de]quinoxaline-6-carboxamide as a pale yellow solid. Physical characteristics:
mp 86-
87°C; 1H-NMR (400MHz, DMSO-d6): 8 10.35, 8.71, 7.76, 7.54, 7.38, 7.20,
7.11,
6.74, 5.56, 5.17, 4.87, 5.56, 3.68, 3.59, 2.99, 2.90, 2.71, 2.29; HRMS (FAB)
found
585.1890 for C32Hz9CLN405 +Hl ; Specific Rotation [a]25D = -7 (c 0.97, DMSO).
Example 35 Preparation of 9-{ [[(2S)-2-(1-benzofuran-2-yl)-2-
hydroxyethyl](methyl)amino]methyl}-N-(4-chlorobenzyl)-1-methyl-2,7-dioxo-2,3-
dihydro-1H,7H-pyrido[1,2,3-de]quinoxaline-6-carboxamide.
0 0
i' I ~ I H I
N' CI
~N\ J
~O
As described in the general procedure outlined in Example 9, 0.56 g of N-(4-
chlorobenzyl)-9-(chloromethyl)-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-
pyrido[1,2,3-
de]quinoxaline-6-carboxamide, 0.50 g of (S~-1-(1-benzofuran-2-yl)-2-
(methylamino)ethanol, 0.46mL of i-Pr2NEt, and 0.65 g of 4A sieves in DMF (35
mL)
affords 0.70 g of 9-{ [[(2S)-2-(1-benzofuran-2-yl)-2-
hydroxyethyl](methyl)amino]
methyl}-N-(4-chlorobenzyl)-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[1,2,3-
de]quinoxaline-6-carboxamide as a pale yellow solid. Physical characteristics:
mp
190-191°C; 1H-NMR (400MHz, DMSO-d6): 8 10.35, 8.71, 7.76, 7.54, 7.38,
7.20,
7.11, 6.74, 5.56, 5.17, 4.87, 5.56, 3.68, 3.59, 2.99, 2.90, 2.71, 2.29; HRMS
(FAB)
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found 585.1912 for C32H29CLN4O5 +Hl ; Specific Rotation [a,]ZSD = 7 (c 0.99,
DMSO).
Example 36 Preparation of 9-{ [[(2R)-2-(1-benzofuran-2-yI)-2-
hydroxyethyl] (methyl)amino]methyl }-N-(4-fluorobenzyl)-1-methyl-2,7-dioxo-2,3-
dihydro-1 H,7H-pyrido [ 1,2, 3-de] quinoxaline-6-carboxamide.
0 0
i' I ~ I H
~N' F
~ N\ J
(~O
to As described in the general procedure outlined in Example 9, 0.28 g of N-(4-
fluorobenzyl)-9-(chloromethyl)-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[
1,2,3-
de]quinoxaline-6-carboxamide, 0.26 g of (R)-1-(1-benzofuran-2-yl)-2-
(methylamino)ethanol, 0.24 mL of i-Pr2NEt, and 0.34 g of 4~ sieves in DMF (30
mL)
affords 0.26 g of 9-{[[(2R)-2-(1-benzofuran-2-yl)-2-
hydroxyethyl](methyl)amino]
15 methyl}-N-(4-fluorobenzyl)-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-
pyrido[1,2,3-
de]quinoxaline-6-carboxamide as a white solid. Physical characteristics: mp 94-
96°C;
1H-NMR (400MHz, CDC13): 8 10.25, 8.63, 7.98, 7.50, 7.15-7.30, 7.01, 6.65,
4.97,
4.95, 4.64, 3.82, 3.70, 3.39, 3.02, 2.87, 2.41; HRMS (ESn found 569.2190 for
C32H29N4~5F' +Hl ; Specific Rotation [a]a5D = -7 (c 0.68, DMSO).
Example 37 Preparation of 9-{[[(2S)-2-(1-benzofuran-2-yl)-2-
hydroxyethyl] (methyl)amino]methyl }-N-(4-fluorobenzyl)-1-methyl-2,7-dioxo-2,3-
dihydro-1 H,7H-pyr ido [ 1,2, 3-de] quinoxaline-6-carboxamide.
0 0
0 off N I ~ I H I ~
~N' F
~N\J
IlO
As described in the general procedure outlined in Example 9, 0.25 g of N-(4-
fluorobenzyl)-9-(chloromethyl)-1-methyl-2,7-dioxo-2, 3-dihydro-1 H,7H-pyrido [
1,2, 3
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de]quinoxaline-6-carboxamide, 0.23 g of (S)-1-(1-benzofuran-2-yl)-2-
(methylamino)ethanol, 0.24 mL of i-Pr2NEt, and 0.30 g of 4A sieves in DMF (30
mL)
affords 0.22 g of 9-{[[(2S)-2-(1-benzofuran-2-yl)-2-
hydroxyethyl](methyl)amino]
methyl }-N-(4-fluorobenzyl)-1-methyl-2,7-dioxo-2,3-dihydro-1H,7H-pyrido [
1,2,3-
de]quinoxaline-6-carboxamide as a pale yellow solid. Physical characteristics:
mp
195-197°C;1H-NMR (400MHz, CDC13): 8 10.25, 8.67, 8.01, 7.52, 7.20-7.40,
7.03,
6.68, 4.90-5.10, 4.65, 3.86, 3.75, 3.45, 3.04, 2.90, 2.45; HRMS (ESI) found
569.2217
for C32H29N4~5F +Hl ; Specific Rotation [a]ZSD = 7 (c 0.93, DMSO).
l0 Preparations of Intermediates:
(1) Preparation of 1-(1-benzofuran-2-yl)-2-(methylamino)ethanol.
KOH (9.2 g), and water (3.7 mL) are added to acetonitrile (125 mL).
Benzofuran-2-carbaldehyde (12.0 g) is dissolved into the acetonitrile mixture.
Trimethylsulfonium iodide (16.7 g) is added and the mixture is stirred at
60°C for 3 h.
The reaction mixture is cooled to room temperature and filtered. The filtrant
is
washed with diethyl ether and filtered. This process is repeated until no more
solid
precipitates. The solution is concentrated iu vacuo and the resulting residue
is
dissolved in a solution of methylamine in methanol (2.0 M, 410 mL). The
mixture is
stirred at room temperature for 18 hours, then concentrated in vacuo to a
brown oil.
The oil is purified by column chromatography (MeOH/CHCl3, 5%, 10%; NH-
40H/MeOH/CHC13, 1:10:89, 1:20:79) to afford 3.0 g of the title compound as an
off
white solid. Physical characteristics: 1H NMR (300 MHz, DMSO-d~ b 7.59, 7.53,
7.24, 6.47, 5.62, 4.77, 2.83, 2.32.
(2) Preparation of 1-(3a,7a-dihydro-1-benzothien-3-yl)-2-(methylamino)ethanol.
Potassium hydroxide (3.11 g) and H20 (0.12 mL) are added to acetonitrile (50
mL). Trimethylsulfonium iodide (5.65 g) and thianaphthene-3-carboxaldehyde
(4.50
g) are then added. The reaction mixture is heated to 60 °C for 4 h. The
reaction
mixture is allowed to cool to room temperature and is then diluted with Et20
(25 mL).
The precipitate is filtered off, and the filtrate is concentrated in vacuo.
The resulting
crude material is dissolved in methanol (40 mL) and added to a solution of
methylamine in methanol (2M, 100 mL). The reaction mixture is stirred at room
temperature for 3 d. The reaction mixture is concentrated ih vacuo. The
resulting
CA 02471089 2004-06-18
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brown oil is purified via column chromatography (CHC13/methanol, 95/5, 90/10;
CHC13/methanol/NH4.OH, 90/10/1) to yield 1.75 g of the title compound as a
yellow
solid. Physical characteristics: Mp 98-102°C; 1H NMR (400 MHz, DMSO-d6)
8 7.98-
7.90, 7.51, 7.60, 7.40-7.33, 5.43, 5.04, 2.80, 2.34; MS (ESI+) m/z 208 (M+H)+.
(3) Preparation of 1-(2,5-dimethyl-3-furyl)-2-(methylamino)ethanol.
3-acetyl-2,5-dimethylfuran (13.3 mL) is dissolved in 1/2 dioxane/Et20 (600
mL) and cooled to 0 °C. Bromine (16.0 g) is added dropwise over 1 h.
The reaction
mixture is stirred at 0 °C for 1 h and then allowed to warm to room
temperature. The
reaction mixture is stirred at room temperature for 18 h. The reaction mixture
is
cooled to 0 °C and an additional 1.0 mL of bromine is added. The
reaction mixture is
allowed to warm to room temperature and is stirred for 2 h. The reaction is
quenced
with a saturated ammonium chloride solution (100 mL). The organic layer is
removed
and the aqueous layer extracted with Et20 (2 x 100 mL). The combined organic
layers are dried (MgS04), filtered, and concentrated iu vacuo. The resulting
brown,
oily solid is purified via column chromatography (hexanes/CH2Cla; 70/30) to
yield a
yellow oily solid which is recrystallized from hexanes to yield 7.52 g of 2-
bromo-1-
(2,5-dimethyl-3-fuiyl)ethanone as a white solid. Physical characteristics: Mp
56-58
°C; 1H NMR (400 MHz, DMSO-d6) 8 6.49, 4.54, 2.50, 2.08; 13C NMR (100
MHz,
2o CDCl3) 8 187.2, 159.3, 150.5, 118.9, 105.6, 33.2, 14.4, 13.2.
2-bromo-1-(2,5-dimethyl-3-furyl)ethanone (7.30 g) is dissolved in methanol
(80 mL) and added dropwise to a solution of methylamine in methanol (2M, 168
mL)
at 0 °C. The reaction mixture is stirred at 0 °C for 30 min and
then sodium
borohydride ( 1.91 g) in H20 (40 mL) is added dropwise. The reaction mixture
is
stirred at 0 °C for 1.5 h and then allowed to warm to room temperature.
The reaction
mixture is stirred at room temperature for 18 h. An additional 0.636 g of
sodium
borohydride is added and stirring is continued for 3 h. The reaction is
quenched with
a 1 N HCl solution and concentrated iu vacuo to remove methanol. The residue
is
poured into cold 2 N NaOH ( 100 mL)/ethyl acetate (200 mL). The organic layer
is
3o removed and the aqueous layer extracted with ethyl acetate (3 x 200 mL).
The
combined organic layers are dried (MgS04), filtered, and concentrated in
vacuo. The
resulting yellow oil is purified via column chromatography (CHC13/methanol,
95/5,
90/10; CHC13/methanol/NH40H, 90/10/1). The resulting pale yellow solid is
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recrystallized from ethyl acetate to yield 2.406 g of the title compound as a
white
solid. Physical characteristics: Mp 76-77 °C; 1H NMR (400 MHz, DMSO-d6)
8 5.93,
4.82, 4.47-4.43, 2.64-2.54, 2.46-2.42, 2.32, 2.16; MS (ESI+) m/z 170 (M+H)+.
(4) Preparation of 1-(methy!amino)-3-phenylpropan-2-ol.
(2,3-epoxypropyl)benzene (5.00 g) is added to a solution of methylamine in
methanol (2.0 M, 187 mL). The reaction mixture is stirred at room temperature
for
18 h and then concentrated in vacuo. The resulting yellow oil is purified via
column
chromatography (CH2C12lmethanol, 95/5, 50/50) to yield 3.89 g of the title
compound
l0 as a yellow oil. Physical characteristics: 1H NMR (400 MHz, DMSO-d6) 8)
7.28-7.15,
4.57, 3.74-3.68, 2.72-2.58, 2.43-2.36, 2.26; MS (ESI+) m/z 166 (M+H)+.
(5) Preparation of 2-(methy!amino)-1-quinolin-2-ylethanol.
Potassium hydroxide (3.21 g) and H20 (0.13 mL) are added to acetonitrile (50
mL). Trimethylsulfonium iodide (5.84 g) and 2-quinoline carboxaldehyde (4.50
g) are
then added. The reaction mixture is heated to 60 °C for 4 h. The
reaction mixture is
allowed to cool to room temperature and is diluted with Et~O (25 rnL). The
precipitate is filtered off. The filtrate is concentrated in vacuo and the
residue is re-
subjected to the reaction conditions above and heated to 60 °C for 1 h.
The reaction
mixture is allowed to cool to room temperature and is diluted with Et20 (25
mL).
The precipitate is filtered off and the filtrate is concentrated ih vacuo. The
resulting
crude material is dissolved in methanol (20 mL) and added to a 2.0 M solution
of
methylamine in methanol (100 mL). The reaction mixture is heated to reflux for
1 h.
The reaction mixture is allowed to cool to room temperature and concentrated
in
vacuo. The resulting brown oil is purified via column chromatography
(CHC13/methanol, 95/5, 90/10; CHC13/methanol/NH40H, 90/10/1) to yield 1.191 g
of
the title compound as a yellow-green oil. Physical characteristics: 1H NMR
(400
MHz, DMSO-d6) ~ 8.36-8.33, 7.98-7.94, 7.76-7.67, 7.59-7.54, 5.63, 4.88-4.84,
2.89-
2.72, 2.32; MS (ESI+) m/z 203 (M+H)+.
(6) Preparation of 2-(methylamino)-1-(5-methyl-2-furyl)ethanol.
Bromine (5.1 mL) is added dropwise over 1 h to a solution of 2-acetyl-5-
methylfuran ( 11.0 g) in dioxane/BtZO ( 1/2, 60 mL) at 0 °C (internal).
The reaction
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mixture is stirred at 0 °C for 30 min and then allowed to warm to room
temperature
and is stirred for 18 h. The reaction mixture is cooled to 0 °C
(internal), and
additional bromine (1.53 mL) is added dropwise. The reaction mixture is
allowed to
warm to room temperature and is stirred for 1 h. A saturated ammonium chloride
solution (100 mL) is added. The organic layer is removed and the aqueous layer
is
extracted with Et20 (2 x 100 mL). The combined organic layers are dried
(MgSO4),
filtered, and concentrated ire vacuo. The resulting brown solid is purified
via column
chromatography (hexanes/CHaCl2, 70/30) to yield a yellow solid which is
recrystallized from EtOAc/hexanes to yield 8.571 g of 2-bromo-1-(5-methyl-2-
to furyl)ethanone as a pale yellow solid. Physical characteristics: Mp 60-63
°C; 1H
NMR (400 MHz, DMSO-d6) 8 7.60, 6.44, 4.58, 2.41.
A solution of 2-bromo-1-(5-methyl-2-furyl)ethanone (8.00 g) in methanol (100
mL) is added dropwise to a 2.0 M solution of methylamine in methanol ( 197 mL)
at 0
°C (internal). The reaction mixture is stirred at 0 °C for 30
min. A solution of sodium
borohydride (2.23 g) in H~,O (40 mL) is then added dropwise. The reaction
mixture is
stirred at 0 °C for 1.5 h and then quenched with a 2 N HCl solution (to
pH 3-4). The
reaction mixture is concentrated i~z vacuo to remove methanol and then poured
into
cold EtOAc (200 rnL)/ 2 N NaOH (100 mL). The organic layer is removed. The
aqueous layer is adjusted to pH 12 with a 2 N NaOH solution and extracted with
EtOAc (3 x 200 mL). The combined organic layers are dried (MgS04), filtered,
and
concentrated irz vacuo. The resulting yellow oil is purified via column
chromatography (CHC13/methanol, 95/5, 90/10; CHC13/methanol/NH40H, 90/10/1).
The resulting yellow oil is crystallized from diethyl ether to yield 1.88 g of
the title
compound as a yellow solid. Physical characteristics: Mp 40-45 °C;1H
NMR (400
MHz, DMSO-d6) b 6.11, 5.97-5.96, 5.05, 4.54-4.51, 2.72-2.65, 2.29, 2.22; MS
(ESI+)
fnlz 156 (M+H)+.
(7) Preparation of 1-(2-Furyl)-2-(methylamino)ethanol.
Bromine (6.5 mL) is added dropwise over 1 h to a solution of 2-acetylfuran
(11.0 g) in dioxane/Et20 (1/2, 60 mL) at 0 °C (internal). The reaction
mixture is then
allowed to warm to room temperature and is stirred for 2 h. A saturated
ammonium
chloride solution (70 mL) is added. The organic layer is removed and the
aqueous
layer is extracted with diethyl ether (2 x 50 mL). The combined organic layers
are
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dried (MgS04), filtered, and concentrated ih vacuo. The resulting brown solid
is
purified by column chromatography (hexanes/CH2Cl2, 70/30) to yield 7.996 g of
2-
furoyl bromide as a yellow oil. Physical characteristics: 1H NMR (400 MHz,
DMSO-
d6) 8 8.09, 7.66-7.64, 6.79-6.77, 4.65.
A solution of 2-furoyl bromide (7.50 g) in methanol (40 mL) is added
dropwise to a 2.0 M solution of methylamine in methanol (198 mL) at 0
°C (internal).
The reaction mixture is stirred at 0 °C for 30 min. A solution of
sodium borohydride
(2.25 g) in water (40 mL) is then added dropwise. The reaction mixture is
stirred at 0
°C for 30 min and then quenched with a 2 N HCl solution (to pH 3-4).
The reaction
l0 mixture is concentrated in vacuo to remove methanol and is then poured into
cold
EtOAc (200 mL)/ 2 N NaOH ( 100 mL). The organic layer is removed. The aqueous
layer is adjusted to pH 12 with a 2 N NaOH solution and extracted with EtOAc
(3 x
200 mL). The combined organic layers are dried (MgS04), filtered, and
concentrated
i~z vacuo. The resulting brown oil is purified by column chromatography
(CHC13/methanol, 95/5; CHC13/methanol/NH40H, 90110/1) to yield 2.06 g of the
title
compound as a brown oil. Physical characteristics: 1H NMR (400 MHz, DMSO-d6) 8
7.56, 6.39-6.37, 6.26-6.25, 5.15, 4.62-4.58, 2.77-2.66, 2.33; MS (ESI+) ~r~lz
142
(M+H)+.
(8) Preparation of I-(3-Furyl)-2-(methylamino)ethanol.
Trimethylsulfonium iodide (20.4 g) and 3-furaldehyde (8.65 mL) are added to
potassium hydroxide (11.2 g) and HBO (0.45 mL) in acetonitrile (150 mL). The
reaction mixture is heated to 60 °C for 2.5 h. The reaction mixture is
allowed to cool
to room temperature. The precipitate is filtered off, and the filtrate is
concentrated iu
vacuo. The resulting crude material (10.747 g) is dissolved in methanol (50
mL) and
added to a 2.0 M solution of methylamine in methanol (100 mL). The reaction
mixture is stirred at room temperature for 3 d and then heated to reflux for
30 min.
The reaction mixture is allowed to cool to room temperature and is
concentrated ii2
vacuo. The resulting brown oil is purified via column chromatography
(CHC13/methanol, 95/5, 90/10; CHC13/methanol/NH4OH, 90/10/1) to yield 2.703 g
of
the title compound as a yellow oil. Physical characteristics: 1H NMR (400 MHz,
DMSO-d6) 8 7.56-7.55, 7.51, 6.44, 5.07, 4.58-4.55, 2.62-2.56, 2.30; MS (ESI+)
m/z
142 (M+H)+.
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(9) Preparation of 2-(methylamino)-1-(6-methylpyridin-2-yl)ethanol.
Potassium hydroxide ( 11.2 g) and H20 (0.45 mL) are added to acetonitrile
(150 mL). Trimethylsulfonium iodide (20.4 g) and 6-methyl-2-pyridine
carboxaldehyde (12.1 g) are then added. The reaction mixture is heated to 60
°C for 3
h. The reaction mixture is allowed to cool to room temperature. The
precipitate is
filtered and the filtrate is concentrated ih vacuo. The resulting crude
material (13.5 g)
is dissolved in methanol (50 mL) and added to a 2.0 M solution of methylamine
in
methanol (250 mL). The reaction mixture is heated to reflux for 30 min. The
reaction
mixture is concentrated in vacuo. The resulting brown oil is purified via
column
chromatography (CHC13/methanol, 95/5, 90/10; CHCl3/methanol/NH40H, 90/10/1).
The resulting brown oil is suspended in hot methanol and the insoluble
material
filtered off. The filtrate is concentrated ih vacuo to yield 3.657 g of the
title
compound as a yellow solid. Physical characteristics: Mp 33-38 °C;1H
NMR (400
MHz, DMSO-d6) 8 7.64, 7.29, 7.10, 5.40, 4.63-4.60, 2.79-2.75, 2.61-2.54, 2.43,
2.30;
MS (ESI+) m/z 167 (M+H)+.
(10) Preparation of 1-(3-methoxyphenyl)-2-(methylamino)ethanol.
A sealed tube containing 2-(3-methoxyphenyl)oxirane (J. Med. Chem. 1992,
35, 3045) (4.0 g) methylamine (27 mL, 2M solution in methanol) are heated for
4 hrs
at 100 °C. After cooling, the reaction is concentrated under reduced
pressure and the
residual oil is purified by silica gel chromatography (Biotage 40M, 6%
methanol/dichloromethane) to yield the title compound (1.1 g) as an off white
solid.
Physical characteristic: 1H NMR (400 MHz, CDC13) 8 7.28, 6.97, 6.81, 4.77,
3.83,
2.99, 2.80, 2.49.
(11) Preparation of 5-[1-Hydroxy-2-(methylamino)ethyl]thiophene-2-
carbonitrile.
To a solution of 2-acetyl-5-cyanothiophene ( 1.5 g) in 20 mL of p-
dioxane/ethyl
ether (1:2, v/v) is added 0.50 mL of bromine. The reaction is stirred at room
temperature for 2 hours. Ice water (30 mL) is added. The resulting solid is
collected
by filtration and is washed with water to yield 1.4 g of 5-
(bromoacetyl)thiophene-2-
carbonitrile as a white solid. The filtrate is allowed to stand overnight to
yield 0.86 g
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of 5-(bromoacetyl)thiophene-2-carbonitrile as a white solid. Physical
characteristics:
1H NMR (DMSO-d6) 8 8.16, 8.11, 4.94; MS (ESI-) m/z 230 (M-H)+.
To a mixture of 5-(bromoacetyl)thiophene-2-carbonitrile (1.85 g) in 50 mL of
methanol at -10 °C is added NaBH4 (0.46 g) in 5 mL of water. After
stirring for 10
minutes, HBr is added to adjust the pH to 3. The reaction mixture is
concentrated to
approximately 25 mL before water (30 mL) is added. The mixture is extracted
with
dichloromethane (3 x 40 mL). The organic phases are combined, washed with
brine,
and dried over MgS04 and concentrated to give 5-(2-bromo-1-
hydroxyethyl)thiophene-2-carbonitrile (1.6 g) as an orange oil. Physical
l0 characteristics: IH NMR (DMSO-d6) 8 7.86, 7.23, 6.67, 5.17, 3.81, 3.68; MS
(ESI-)
mlz 232 (M-H)+.
To a solution of 5-(2-bromo-1-hydroxyethyl)thiophene-2-carbonitrile (1.6 g) in
20 mL of methanol is added 80 mL of methylamine solution (2.0 M in methanol).
The reaction mixture is stirred at room temperature overnight. The reaction is
concentrated and the resulting residue is dissolved in 20 mL of methanol and
treated
with BioRad AG" 50w-x2 resin (2 g, hydrogen form, strongly acidic ration) for
4
hours. The resin is collected by filtration and is washed with methanol. The
resin-
bound product is eluted off with 10% NH40H/MeOH ( 100 mL). The ammonium
hydroxide filtrate is concentrated under reduced pressure. The residue is
flash column
chromatographed on silica gel eluting with 1% NH40H/10% MeOH/89% CH2C12 to
yield 0.80 g of the title compound as a white solid. Physical characteristics:
1H
NMR (DMSO-d6) S 7.81, 7.13, 6.13, 4.93, 2.72, 2.33; MS (ESI+) m/z 183 (M+H)+;
HRMS found 183.0600.
(12) Preparation of 2-(methylamino)-1-pyrimidin-2-ylethanol.
2-Acetylpyrimidine (Chimia 1996, 50, 538 and J. Het. Ch.em. 1994, 31, 1041)
(7.37 g) and diisopropylethylamine (23.4 g) are dissolved in dry CH2C12 under
N2 then
cooled in an ice bath. Triisopropylsilyl trifluoromethanesulfonate (17.9 mL)
is added
over 2-3 min and the reaction is stirred overnight. The solvent is removed
under
reduced pressure and the residue is treated with ether (200 mL), filtered and
washed
with saturated sodium bicarbonate solution (2 x 50 mL). Evaporation of the
solvent
gives 2-{ 1-[(triisopropylsilyl)oxy]vinyl}pyrimidine as a red oil. Physical
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characteristics: HRMS (FAB) found 279.1898 (M+H)+; 1H NMR (300 MHz, CDCl3)
8 1.15, 1.31, 4.90, 5.82, 7.16, 8.74.
N-chlorosuccinimide (9.97 g) is added to a solution of { 1-
[(triisopropylsilyl)oxy]vinyl}pyrimidine (17.3 g) in dry THF (120 mL) under N~
then
heated at 65~~ for 5 h. After cooling, ether (275 mL) is added and then washed
with
saturated sodium bicarbonate solution (2 x 100 mL). The organic layer is dried
over
sodium sulfate, filtered and concentrated under reduced pressure. The
resulting oil is
dissolved in hexanes (250 mL), treated with MgS04 and filtered. Concentration
at
reduced pressure affords 2-{2-chloro-1-
[(triisopropylsilyl)oxy]ethenyl}pyrimidine as a
l0 yellow oil. Physical characteristics: HRMS (FAB) found 313.1509(M+H)+; iH
NMR
(300 MHz, CDC13) 8 1.13, 1.33, 6.97, 7.17, 8.68.
2-{2-Chloro-1-[(triisopropylsilyl)oxy]ethenyl}pyrimidine (19.4 g) is dissolved
in acetonitrile (90 mL) and treated with 48% HF (10 mL) for 4 h. Saturated
sodium
bicarbonate solution (ca. 250 mL) is then added carefully to adjust the pH to
~ 7 and
the mixture is extracted with CHaCl2 (3 x 200 mL). The combined organic layers
are
dried (Na2S04), filtered and concentrated. The resulting biphasic mixture is
decanted
to remove the upper phase and the lower oil is chromatographed over silica gel
eluting
with 2.5% MeOH-CHC13 to give 6.5 g of 2-chloro-1-pyrimidin-2-ylethanone as a
pale
yellow solid. Physical characteristics: mp 73-80°C; Anal. Found: C,
46.05; H, 3.09;
N, 17.93.
2-Chloro-1-pyrimidin-2-ylethanone (6.15 g) is dissolved in ethanol (125 mL)
and CeC13.7H20 (14.64 g) is added. The mixture is stirred for 10 min, then
sodium
borohydride (1.49 g) is added over 2 min. After lhr, the solid is filtered and
the filtrate
evaporated. The resulting residue is treated with saturated ammonium chloride
solution (25 mL), followed by brine (250 mL) and the mixture adjusted to pH 3-
4 with
1N HCl. Extraction with ethyl acetate (3 x 250 mL) followed by concentration
at
reduced pressure affords an oil which is chromatographed over silica gel to
give 3.85
g of 2-chloro-1-pyrimidin-2-ylethanol as a pale yellow oil. Physical
characteristics: IR
(liq.) 3427, 2404, 2346, 2196, 1980, 1568, 1439, 1425, 1390, 1183, 1111, 1087,
812,
658, 636 cm -1.
In a pressure bottle is placed 2-chloro-1-pyrimidin-2-ylethanol (3.525 g),
sodium iodide (0.344 g) and a 2M solution of methylamine in methanol (160 mL).
The bottle is sealed and heated at 62°C for 17 h. The solvent is
evaporated and the
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residue is stirred with 10% MeOH-CHC13. The mixture is filtered and
concentrated to
give an oil that is chromatographed over silica gel eluting with 5-10% MeOH-
CH2C12
containing 1 % triethylamine. The product containing fractions are combined
and
concentrated to give the title compound as an amber oil (1.625 g). Physical
characteristics: 1H NMR (400 MHz, CDC13) 8 2.53, 3.03, 3.21, 3.66, 5.03, 7.26,
8.77;
HRMS (FAB) found 263.0636 (M+H)+.
(13) Preparation of R-2-(1-hydroxy-2-N-methylamino-ethyl)-pyridine.
2-Acetylpyridine (50 g) is placed in a 2L 1N round bottom flask and
to anhydrous CH2C12 (Alrdich Sure Seal, 0.65 L) is added, followed by the
addition of
i-Pr2NEt (160.27 g). The flask is equipped with a 125 mL pressure equalized
dropping
funnel, and the mixture is placed under nitrogen and cooled in an ice-water
bath. To
the chilled ketone/amine mixture is added TIPSOTf (139.7 g) over 1.5 hours.
The
mixture is allowed to warm to room temperature overnight. The reaction mixture
is
concentrated ih vacuo on a rotary evaporator (T<_ 25°C) to give a
yellow oil and a
white solid. The flask contents are transferred to a 2L separatory funnel with
ether
(1.2L) resulting in the formation of additional white solid material and the
mixture i.s
washed with saturated aq. NaHC03 (2 x 0.65L). The organic phase is separated,
'dried
over NaaS04, then is concentrated in vacuo to furnish the crude 2-[1-Tri-
isopropylsilyloxy-vinyl]-pyridine (131.5g) as a yellow-orange oil. This crude
material
is not further purified, but is immediately carried to the next step. 1H-NMR
(400MHz,
CDC13) b = 8.57, 7.71, 7.21, 5.65, 4.58, 1.36, 1.15. '
Crude 2-[1-Tri-isopropylsilyloxy-vinyl]-pyridine (131.5g) is placed in a 2L,
1N round bottom flask and dissolved in anhydrous THF (Aldrich Sure Seal,
0.6L).
The flask is equipped with a reflux condenser and the apparatus is placed
under
nitrogen. NCS (60.66g) is added and the mixture is heated to reflux and
maintained at
reflux for 2 hours. The reaction mixture is cooled to room temperature, poured
into a
4L separatory funnel containing ether (1.5L), and is washed with saturated aq.
NaHCO3 (2 x 0.7L). The organic phase is separated, dried (Na2S04), and
concentrated
in vacuo to afford crude 2-[1-tri-isopropylsilyloxy-2-chloro-vinyl]-pyridine
(117.5 g)
as a yellow-orange oil. The crude material is not further purified, but is
immediately
carried into the next step. 1H-NMR (400MHz, CDCl3) b 8.53, 7.71, 7.52, 7.22,
6.58,
1.21, 1.13.
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Crude 2-[1-tri-isopropylsilyloxy-2-chloro-vinyl]-pyridine (117.3 g) is placed
in
a 4L plastic bottle and is dissolved in acetonitrile (0.4 L). To the stirring
solution is
added 48% aqueous HF (170 mL) and the progress of the reaction is monitored by
reverse phase analytical HPLC. After ~ 2 hours the reaction is judged to be
complete,
and the pH of the solution is carefully adjusted to ca. 8 with saturated aq.
NaHC03.
The mixture is poured into a separatory funnel containing CH2C12 (1.5L). The
organic
phase is removed and the aq. Layer is extracted with CH2C12 (2 x 1.OL). The
combined organic layers are dried (Na2S04), and concentration in vacuo to give
the
crude 2-chloroacetyl pyridine (49.5 g) as a tan solid after cooling. The crude
material
is used as is in the asymmetric reduction. 1H-NMR (400MHz, CDC13) 8 8.66,
8.09,
7.88, 7.54, 5.12.
[RuCl2(r~6 p-cymene)]2 (0.84g), Et3N (0.67g), and (IR, 2R)-N p-
toluenesulfonyl-1,2-diphenylethylenediamine (l.Og) are combined in a 500mL 1N
round bottom flask. i-PrOH is added, a reflex condenser is attached and the
mixture is
warmed under reflex, and maintained, for 1 hour. The mixture is cooled to room
temperature and is concentrated in vacuo (rotovapor followed by vacuum pump)
to
furnish the catalyst as a brown powdery solid. To the catalyst is added
anhydrous
DMF (Aldrich Sure Seal, 225mL), followed in order by 2-chloroacetylpyridine
(23.88g) and HCOOH/Et3N (5:2, Fluka, 55mL). Reaction progress is monitored by
reverse phase analytical HPLC, and after 65 minutes of stirring, the starting
material
had been consumed. Quench the reaction by adding MeOH (25mL), stir 5 minutes
and
then the solvent is removed iia vacuo (cold finger rotovapor, vacuum pump) to
give a
red-black viscous oil. The crude material is taken up in Et20lCH2C12 (4: I,
1.25L),
placed in a 3L separatory funnel, washed with saturated aq. NaHC03 (I.OL),
brine
(1.OL), and dried (Na2SO4). Filtration and concentration in vacuo afforded the
crude
product as a red-orange oil which is purified by chromatography on a column of
silica
gel (70mm OD, 250g 230-400mesh, packed hexanes; compound applied in
CHZCIa/hexanes 60:40; eluted with hexanes/Et20 (75:25; 65:35; 55:45) using the
flash technique. Product fractions are combined to afford 16.41g of the target
5-2-(1-
hydroxy-2-chloroethyl)-pyridine as pale yellow solid. mp 49-50°C; 1H-
NMR
(400MHz, CDCl3): 8 8.60, 7.77, 7.58, 7.30, 5.00, 4.20, 3.85; Anal. Found: C,
53.27;
H, 5.19; N, 8.81, Cl, 22.29; Specific Rotation [a]DDS = 62 (c 0.94, methanol).
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S-2-(1-hydroxy-2-chloroethyl)-pyridine (6.Og) and NaI (0.57g) are combined
in a 500mL, plastic coated, thick walled bottle and are covered with 2M MeNH2
in
MeOH (0.19L). The Teflon stopper is wrapped in Teflon tape and the bottle is
sealed.
Stirring is started, and the bottle is immersed in a 60°C oil bath for
16 hours. The
yellow-brown mixture is cooled to room temperature. Concentration in, vacuo
affords
the crude product as a yellow oil, which is treated with CHZCh-THF (0.25L,
10:90) to
give a yellow solution and a white precipitate. The precipitate is removed by
filtration,
is rinsed with CH2C12-THF ( 10:90) and the combined filtrated are concentrated
in
vacuo to give a yellow-brown oil. The crude product is purified by
chromatography on
to a column of silica gel (70mm OD, 250g, 230-400mesh; packed with CH2C12-MeOH
90:10; eluted with CH2Cl2-MeOH 90:10, CHZC12-MeOH-NH4OH 89:10:1) using the
flash technique. Product fractions are combined to provide 3.18g of
aminoethanol R-
2-(1-hydroxy-2-N-methylamino-ethyl)-pyridine as an amber oil. 1H-NMR (400MHz,
DMSO-d6): 8 = 8.49, 7.79, 7.52, 7.25, 4.75, 2.90, 2.67, 2.32; HRMS (FAB):
Found
153.1009; Specific Rotation [a]Das = 49 (c 0.36, CHZC12).
(14) Preparation of N-Methyl R-1-(2-furyl)-2-aminoethanol.
2-Acetylfuran (50g) is placed in a 2L 1N round bottom flask and anhydrous
CH2C12 (Alrdich Sure Seal, 0.70L) is added, followed by the addition of i-
Pr2NEt
(176g). The flask is equipped with a 125mL pressure equalized dropping funnel,
and
the mixture is placed under nitrogen and cooled in an ice-water bath. To the
chilled
ketone/amine mixture is added TIPSOTf (153.2g) over 1.5 hours. The mixture is
allowed to warm to room temperature overnight. The reaction mixture is
concentrated
ih vacuo on a rotary evaporator (T<_ 25°C) to give a yellow oil and a
white solid. The
flask contents are transferred to a 2L separatory funnel with ether (1.2L)
resulting in
the formation of additional white solid material and the mixture is washed
with
saturated aq. NaHC03 (2 x 0.70L). The organic phase is separated, dried over
Na2S04,
then is concentrated irc vacuo to furnish the crude 2-[1-tri-isopropylsilyloxy-
vinyl]-
furan (118.3g) as a yellow-orange oil. This crude material is not further
purified, but is
3o immediately carried to the next step. 1H-NMR (400MHz, CDC13) 8 7.36, 6.49,
6.40,
4.86, 4.37, 1.32, 1.14.
Crude 2-[1-tri-isopropylsilyloxy-vinyl]-furan (116.3g1) is placed in a 2L, 1N
round bottom flask and dissolved in anhydrous THF (Aldrich Sure Seal, 0.6L).
The
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flask is placed under nitrogen, cooled in a -10°C bath, then NCS (64.1
lg) is added and
the mixture is stirred for 1 hour, after which time the reaction is judged to
be complete
by analytical reverse phase HPLC. The reaction mixture is warmed to room
temperature, poured into a 4L separatory funnel containing ether (1.5L), and
is washed
with saturated aq. NaHC03 (2 x 0.7L). The organic phase is separated, dried
(Na2S04), and concentrated ih vacuo to afford 2-[1-tri-isopropylsilyloxy-2-
chloro-
vinyl]-furan (129.9g) as a yellow-orange oil. The crude material is not
further purified,
but is immediately carried into the next step. 1H-NMR (400MHz, CDC13) 8 7.36,
6.43,
6.40, 5.95, 1.30, 1.11.
2-[1-Tri-isopropylsilyloxy-2-chloro-vinyl]-furan (129.9g) is placed in a 4L
plastic bottle and is dissolved in acetonitrile (0.6L). To the stirring
solution is added
48% aqueous HF (65mL) and the progress of the reaction is monitored by reverse
phase analytical HPLC. After. Ca. 2 hours the reaction is judged to be
complete, and
the pH of the solution is carefully adjusted to ca. 7 with saturated aq.
NaHC03. The
mixture is poured into a separatory funnel containing CH2C12 (1.5L). The
organic
phase is removed and the aq. layer is extracted with CH2C12 (2X1.OL). The
combined
organic layers are dried (Na~S04), and concentration in vacuo afforded the
crude 2-
chloroacetyl furan (41.9g) as a yellow oil. The crude material is used as is
in the
asymmetric reduction. 1H-NMR (400MHz, CDC13): 8 7.58, 7.33, 6.59, 4.57; MS
(ES+): 145.4 (M+H+).
[RuCl2(r~6 p-cymene)]2 (0.99g), Et3N (0.67g), and (1R, 2R)-N p-
toluenesulfonyl-1,2-diphenylethylenediamine (1.18g) are combined in a 500mL 1N
round bottom flask. i-PrOH (25 mL) and Et3N (0.67g) are added, a reflux
condenser is
attached and the mixture is warmed under reflux, and maintained, for 1 hour.
Cool to
room temperature and concentrate i~c vacuo (rotovapor) to furnish the catalyst
as an
orange-brown powdery solid. To the catalyst is added anhydrous DMF (Aldrich
Sure
Seal, 250mL), followed in order by 2-chloroacetylfuran (20.6g) and HCOOH/Et3N
(5:2, Fluka, 5lmL). Reaction progress is monitored by reverse phase analytical
HPLC,
and after 65 minutes of stirring, the starting material had been consumed.
Quench the
reaction by adding MeOH (25mL), stir 5 minutes and then the reaction mixture
is
poured into ice-water (1L) and the aqueous phase is saturated with salt. The
mixture is
transferred to a 2L separatory funnel with ether (500mL), shaken, and the
organic
phase is removed. The aqueous layer is extracted with ether (3X250mL) and the
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combined organic layers are washed with saturated aq. NaHC03 (0.5L), brine
(4X250mL), and dried (Na2SO4). Filtration and concentration irc vacuo affords
the
crude product as a red-orange oil (20.5g) that is triturated with
ether/pentane (10:90,
4X100mL). The combined triturates are concentrated in vacuo carefully (the
halohydrin is volatile, hence the choice of ether/pentane as triturant and no
removal of
DMF in vacuo) to furnish the desired halohydrin S-1-(2-furyl)-2-chloroethanol
(15.97g). 1H-NMR (400MHz, CDC13) 8 7.41, 6.37, 4.95, 3.85, 2.58; HRMS (E~
Found 146.0136; Specific Rotation [a,]Das = 17 (c 0.97, methanol).
To (S)- 1=(2-furyl)-2-chloroethanol (S.Og) in dry CH2C12 (Aldrich Sure
Seal, 75mL), cooled in an ice-water bath under nitrogen, is added Et3N
(1.38g). Stir
5 minutes, then methyl isocyanate (3.32g) is added via syringe over 2 minutes.
Allow
the ice to melt and the mixture to warm toward room temperature while
monitoring
the reaction by HPLC. Allow to stir overnight and the mixture is poured into
Et20
(0.3L) and brine (0.3L). The organic phase is reserved, the aq. layer is
extracted with
Et20 (2X0.2L); the combined organic phases are washed with brine (0.4L), and
dried
(NaaS04). Concentration ih vacuo afforded the crude carbamate as a brown,
viscous
oil, which is purified by chromatography (Biotage~ 40g column, EtOAc/hexanes
10:90, EtOAc/hexanes 20:80). Product fractions afford 4.56g of S-1-(2-furyl)-2-
chloroethanol-N-methylcarbamate as a clear, pale yellow oil which solidified
to an
2o ivory solid upon cooling. mp 26-27°C; 1H-NMR (400MHz, CDC13) 8 7.43,
6.45, 6.39,
5.97, 4.79, 3'.89, 2.82; 13C-NMR (100MHz, CDC13):8 156.2, 150.3, 143.3, 110.8,
109.9, 69.1, 44.0, 28.0; I~F Moisture: 0.13%; Anal. Found: C, 46.99; H, 4.89;
N, 6.85;
Cl, 17.31; Specific Rotation [a]Das = 94 (c 1.02, CH2C12).
The crude carbamate is dissolved in dry THF (0.2L, Aldrich Sure Seal) and
the solution is cooled in an ice-water bath under nitrogen. To the chilled
carbamate
solution is added KOtBu (1.OM in THF, 97mL) over 15 minutes. The mixture is
allowed to stir after the addition is complete and HPLC analysis suggests that
the
reaction is complete within 15 minutes. The mixture is cast into Et20 (1.25L)
and
brine (1.OL) containing 1N aq. HCL (50mL). The organic phase is separated, the
aqueous layer is extracted with Et20 (1.OL). The combined organic phases are
washed
with saturated aq. NaHC03 (l.OL) and dried (Na2S04). Concentration in vacuo
afforded the crude oxazolidinone as a red-black oil which is triturated with
pentane-
Et20 (2:1; 3 X 0.2L). The pentane-Et20 aliquots are concentrated in vacuo to
give a
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red solid which is purified by chromatography on a 120g Biotage~ column
(introduced as a solution in CH~,C12, eluted with EtOAc/hexanes, 35:65;
EtOAc/hexanes 50:50). Product fractions are combined to afford 8.75g of 5R-3-
methyl-5-(2-furyl)-2-oxazoldinone as a pale yellow oil, which solidified to
furnish an
ivory solid upon cooling. mp 54-55°C; 1H-NMR (400MHz, CDC13) 8 7.47,
6.49, 6.41,
5.46, 3.78, 2.97; 13C-NMR (100MHz, CDC13) S 155.9, 148.1, 142.1, 109.0, 108.4,
65.9, 48.8, 29.4; KF Moisture: 0.07%; Anal. Found: C, 57.46; H, 5.39; N, 8.36;
Specific Rotation [a]DZS = -106 (c 1.01, CH2C12).
To 5R-3-methyl-5-(2-furyl)-2-oxazoldinone (B.Og) in a 500mL 1N RB flask
is added 1N aq. KOH (240mL). The flask is equipped with a reflux condenser,
placed
under nitrogen, then is immersed in a preheated (50°C) oil bath. The
mixture is
allowed to stir and the 5R-3-methyl-5-(2-furyl)-2-oxazoldinone suspension
slowly
affords a clear solution. After stirring for 3 hours at 50°C, HPLC
analysis indicated
that the reaction is complete. The mixture is cooled to room temperature and
is cast
into a separatory funnel, the flask is rinsed into the separatory funnel with
Et~O/CH2Cl2 (95:5, 0.5L) and the aq. layer is saturated with salt. The organic
phase is
removed, the aq. phase is extracted with Et20/CHZC12 (95:5, 2 X 0.5L) and the
combined organic phases are dried (Na2S04). Concentration in vacuo affords N-
methyl R-1-(2-furyl)-2-aminoethanol (6.50g) as a pale orange oil which
solidifies at
2o freezer (-20°C) temperatures. 1H-NMR (400MHz, DMSO-d6) b 7.55, 6.37,
6.25,
4.59, 2.70, 2.25; 13C-NMR (100MHz, DMSO-d6) S 157.3, 141.9, 110.5, 105.9,
65.5,
56.5, 36.5; KF Moisture: 0.83%; Anal. Found: C, 59.90; H, 7.83; N, 9.68;
Specific
Rotation [a]Da5 = 32 (c 0.96, EtOH).
(15) Preparation of S-3-(1-hydroxy-2-N-methylamino-ethyl)-pyridine.
As described for the preparation of N-methyl R-1-(2-furyl)-2-aminoethanol,
3-chloroacetylpyridine (0.075g) (Claervi. Ber. 1951, 84, 147-149) is reduced
with
([RuClz(r~6 p-cymene)]2 (3mg), Et3N (3~,L), and (1R, 2R)-N p-toluenesulfonyl-
1,2-
diphenylethylenediamine (3mg), in DMF (2 mL) containing HCOOH/Et3N (5:2, 0.18
3o mL) to give S-2-( 1-hydroxy-2-chloroethyl)-pyridine (0.055g) as a pale
yellow solid.
1H NMR (300 MHz, CDCl3) 8 8.53, 7.79, 7.32, 4.97, 3.88, 3.72; Chiral HPLC
Analysis (Chiracel OJ): 99:1.
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As described for the preparation of N-methyl R-1-(2-furyl)-2-aminoethanol,
S-2-(1-hydroxy-2-chloroethyl)-pyridine is treated with methyl isocyanate
(3.21mL)
and Et3N (1.76mL) in anhydrous CH~C12 (75mL, Aldrich Sure Seal) to give 5R-3-
methyl-5-(3-pyridyl)-2-oxazoldinone (6.05g) as a white solid. Mp 79-
80°C; 1H-NMR
(400MHz, CDCl3) 8 8.65, 8.59, 7.69, 7.32, 5.93, 5.16, 3.79, 2.81; Specific
Rotation
[a]25D = 33° (c 0.96, chloroform); Anal. Calcd for C9H11C1N2O2: C,
50.36; H, 5.16;
N, 13.05; Cl, 16.52; Found: C, 50.29; H, 5.18; N, 12.96; Chiral HPLC Analysis
(Chiracel OJ): 98.3:1.7.
As described for the preparation of N-methyl R-1-(2-furyl)-2-aminoethanol,
l0 5R-3-methyl-5-(3-pyridyl)-2-oxazoldinone (4.19g) is treated with 1N aq.
I~OH
(0.12L) and THF (50mL) to give S-3-(1-hydroxy-2-N-methylamino-ethyl)-pyridine
(2.36g) as a pale yellow amorphous solid. IR (diffuse reflectance) 3303, 3295,
3087,
3053, 3035, 2977, 2889, 2840, 2793, 2311, 2265, 2178, 2114, 2092, 713crri 1;
HRMS
(ESn found 153.1017 for C8H12N20 +Hl ; Specific Rotation [a]25D = 70°
(c 1.03,
15 methylene chloride); Chiral HPLC Analysis (Chiracel OJ): 98.7:1.3.
(16) Preparation of S-3-(1-hydroxy-2-N-methylamino-ethyl)-thiophene.
As described for the preparation of N-methyl R-1-(2-furyl)-2-aminoethanol,
3-acetylthiophene (50 g) in anhydrous CHZCl2 (Alrdich Sure Seal, 600 mL) is
20 treated with i-Pr2NEt (1.18 mol), T1PSOTf (117.2 mL) to give 3-[1-tri-
isopropylsilyloxy-vinyl]-thiophene (117 g) as a brown oil. This crude material
is not
further purified, but is immediately carried to the next step. 1H-NMR (300MHz,
CDC13) 8 7.26, 7.20, 6.99, 4.79, 4.33, 1.21-1.41, 1.20.
As described for the preparation of N-methyl R-1-(2-furyl)-2-aminoethanol,
25 3-[1-tri-isopropylsilyloxy-vinyl]-thiophene (117 g) is treated with NCS
(52.87g) to
give 1388 of crude 3-[1-tri-isopropylsilyloxy-2-chloro-vinyl]-thiophene. 1H-
NMR
(400MHz, CDCl3) ~ 7.00-7.40, 5.80-5.95, 1.00-1.40.
As described for the preparation of N-methyl R-1-(2-furyl)-2-aminoethanol,
3-[1-tri-isopropylsilyloxy-2-chloro-vinyl]-thiophene (138g) is treated with
48% aq.
3o HF (159mL) in acetonitrile (0.5L) to give 3-chloroacetyl-thiophene (54.6g)
as an
amorphous pale yellow solid. 1H-NMR (400MHz, CDCl3) 8 8.18, 7.60, 7.40, 4.61.
As described for the preparation of N-methyl R-1-(2-furyl)-2-aminoethanol,
3-chloroacetyl-thiophene (43g) is reduced ([RuCl2(r~6 p-cymene)]2 (1.64 g),
Et3N (1.4
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mL), (1R, 2R)-Np-toluenesulfonyl-1,2-diphenylethylenediamine (1.96 g), i-
propanol
(30 mL), anhydrous DMF (Aldrich Sure Seal°, 200 mL), HCOOH/Et3N (5:2,
Fluka,
97 mL) ) to give S-1-(3-thienyl)-2-chloroethanol (27.4 g) as a pale yellow
oil. 1H-
NMR (400MHz, CDC13) 8 7.28-7.40, 7.11, 5.02, 3.82, 3.71; HRMS (En found
161.9905 for C6H7CLOS; Specific Rotation [a]25D = 40° (c 1.01,
methylene
chloride); Chiral HPLC Analysis (Chiracel OJ): 98.8:1.2.
As described for the preparation of N-methyl R-1-(2-furyl)-2-aminoethanol,
S-1-(3-thienyl)-2-chloroethanol (S.OOg) is treated with Et3N (1.24g) and
methyl
isocyanate -(2.98g) in CHZC12 (35mL) to give S-1-(3-thienyl)-2-chloroethanol-N-
l0 methylcarbamate 5.91g as a clear, colorless oil. 1H-NMR (300MHz, CDC13) 8
7.30-
7.40, 7.09, 6.03, 4.82, 3.82), 2.82; Specific Rotation [a]25D = 57° (c
0.73, methylene
chloride); Chiral HPLC Analysis (Chiracel OJ): 98.5:1.5.
As described for the preparation of N-methyl R-1-(2-furyl)-2-aminoethanol,
S-1-(3-thienyl)-2-chloroethanol-N-methylcarbamate (5.39g, in THF (30mL), is
treated
15 with NaH (1.19g) to afford 5R-3-methyl-5-(3-thienyl)-2-oxazoldinone (3.80g)
as a
pale yellow solid. mp 68-69°C; IR (diffuse reflectance) 3096, 2483,
2408, 2350,
2328, 2253, 1755, 1733, 1501, 1439, 1408, 1264, 1249, 1137, 1030 cm 1; HRMS
(ESA found 184.0432 for C$H9NO2S +Hl; Specific Rotation [a]25D = -14°
(c 1.05,
chloroform); Anal. Calcd for C8H9NO~S: C, 52.44; H, 4.95; N, 7.64; S, 17.50.
Found:
20 C, 52.38; H, 5.05; N, 7.60; S, 17.33; Chiral HPLC Analysis (Chiracel OJ):
98.4:1.6.
As described for the preparation of N-methyl R-1-(2-furyl)-2-aminoethanol,
5S-3-methyl-5-(3-thienyl)-2-oxazoldinone (3.35g) is treated with 1N aq. KOH
(95mL)
to provide S-3-(1-hydroxy-2-N-methylamino-ethyl)-thiophene (2.79g) as a pale
yellow
oil. 1H-NMR (300MHz, CDC13) 8 7.31, 7.24, 7.07, 4.86, 2.60-3.00, 2.47; IR
(liq.)
25 3315, 3102, 2972, 2941, 2890, 2857, 2800, 1996, 1473, 1451, 1066, 853, 836,
787,
652 cm 1; HRMS (ESA found 158.0628 for C7H11NOS +Hl ; Specific Rotation [a]25D
= 48° (c 0.86, chloroform); Chiral HPLC Analysis (Chiracel OJ): >99:1.
(17) Preparation of R-2-(1-hydroxy-2-N-methylamino-ethyl)-pyrazine.
30 As described for the preparation of N-methyl R-1-(2-furyl)-2-aminoethanol,
2-acetylpyrazine (53.9 g) in anhydrous CHZC12 (Alrdich Sure Seal, 700 mL) is
treated with i-PrZNEt (171.0 g), TIPSOTf (148.6 g) to give 2-[1-tri-
isopropylsilyloxy-
vinyl]-pyrazine ( 132.9 g) as a brown oil. This crude material is not further
purified,
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but is immediately carried to the next step. 1H NMR (300 MHz, CDC13) 8 8.97,
8.49,
5.66, 4.65, 1.36, 1.14; 13C NMR (100 MHz, CDC13) 8 153.0, 150.0, 143.8, 143.5,
141.1, 94.1, 18.1, 12.7; HRMS (FAB) found 279.1891 for ClSHasNaOSi +Hl.
As described for the preparation of N-methyl R-1-(2-furyl)-2-aminoethanol,
2-[ 1-tri-isopropylsilyloxy-vinyl]-pyrazine ( 132.9 g) in anhydrous THF
(Aldrich Sure
Seal, 640 mL) is chlorinated with NCS (64.78 g) to give 2-[1-tri-
isopropylsilyloxy-2-
chloro-vinyl]-pyrazine ( 169.45 g) as a brown oil. The crude material is not
further
purified, but is immediately carried into the next step. 1H NMR (400 MHz,
CDC13) 8
8.82, 8.51, 8.49, 6.62, 1.33, 1.13; HRMS (FAB) found 313.1511 for
C15H~,SCINaOSi
to +Hl.
As described for the preparation of N-methyl R-1-(2-furyl)-2-aminoethanol,
2-[1-tri-isopropylsilyloxy-2-chloro-vinyl]-pyrazine (169.45 g) is placed in a
4 L
plastic bottle and dissolved in acetonitrile (470 mL) and treated with 48%
aqueous HF
(73.54 mL) to give 2-chloroacetylpyrazine (60.1 g) as a very light yellow
solid. mp
82.6 - 83.8 °C (dec.); IR (diffuse reflectance) 2944, 1716, 1400, 1390,
1318, 1226,
1170, 1163, 1053, 1019, 1000, 851, 807, 790, 685 cm 1; 1H NMR (300 MHz, CDC13)
8 9.23, 8.80, 8.64, 5.01; 13C NMR (75 MHz, CDC13) S 191.4, 148.4, 145.6,
143.5,
143.3, 46.4.
As described for the preparation of N-mthyl R-1-(2-furyl)-2-aminoethanol,
2-chloroacetylpyrazine (60.1 g) is reduced ([RuCl2(r~6 p-cymene)]2 (0.980 g),
Et3N
(0.664 g), (1R, 2R)-Np-toluenesulfonyl-1,2-diphenylethylenediamine (1.17 g) i-
propanol (24.7 mL), anhydrous DMF (Aldrich Sure Seal°, 247 mL),
HCOOH/Et3N
5:2 Fluka, 57.3 mL to ive 1S -2-chloro-1 azin-2- lethanol 22.5 as a
( ~ ) ) g ( ) -pYr Y ( g)
brown oil. IR (liq.) 3275, 3064, 2959, 2869, 1474, 1405, 1308, 1154, 1091,
1059,
1019, 856, 775, 663, 649 cm 1; 1H NMR (400 MHz, CDCl3) 8 8.80, 8.56, 5.07,
3.93,
3.87; 13C NMR (100 MHz, CDC13) ~ 154.2, 144.2, 143.5, 143.3, 71.7, 48.9; HRMS
(FAB) calcd for C6H7C1N20 +Hl 159.0325, found 159.0323; Specific Rotation
~o~]D
+ 40 (c 0.61, ethanol); Anal. Found: C, 45.23; H, 4.60; N, 17.38.
As described for the preparation of R-2-(1-hydroxy-2-N-methylamino-ethyl)-
pyridine, (1S)-2-chloro-1-pyrazin-2-ylethanol (11.8 g), NaI (1.12g) and 2M
MeNH~
in MeOH (370 mL) affords of R-2-(1-hydroxy-2-N-methylamino-ethyl)-pyrazine
(8.18
g) as a light orange liquid that solidified on standing. Mp 78 - 81 °C;
1H NMR (400
MHz, CD3OD) 8 8.79, 8.58, 8.53, 5.00, 3.15, 3.05, 2.55; 13C NMR (75 MHz,
CD30D)
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8 159.1, 145.1, 144.6, 143.9, 71.5, 57.5, 35.6; HRMS (FAB) found 154.0973 for
C7H11N30 +Hl; Specific Rotation ~oc]~ +58 (c 1.02, methanol).
Carbonyldiimidazole (4.26 g) is dissolved in dichloromethane (80 mL). To
this solution is slowly added, via cannula addition, R-2-(1-hydroxy-2-N-
methylamino-
ethyl)-pyrazine (3.66 g) dissolved in dichloromethane (60 mL). The reaction is
stirred
at room temperature for 16 h. The solvent in vacuo and purification is
accomplished
by silica gel column chromatography (98 : 2 dichloromethane - methanol, sample
and
silica gel loaded in dichloromethane). Any uncyclized carbamate collected off
the
column is dissolved in methanol, to which is added a catalytic amount of 1 M
NaOH
and the solution is refluxed until completely cyclized. This freshly cyclized
material
is then purified by column chromatography as above. The combined pure
oxazolidinone fractions are then concentrated iya vacuo to provide (5R)-3-
methyl-5-
pyrazin-2-yl-1,3-oxazolidin-2-one (3.77 g) as a white solid. This material is
upgraded
by chiral preparative HPLC to give material with > 95% ee. Mp 113.5 -114.1
°C; iH
NMR (400 MHz, CDCl3) 8 8.84, 8.62, 8.58, 5.62, 4.02, 3.80, 2.94; 13C NMR (100
MHz, CDC13) 8 157.4, 153.2, 144.8, 144.1, 142.7, 72.2, 51.6, 31.1; HRMS (FAB)
found 180.0781 for C$H9N30~ +Hl ; Specific Rotation ~a]D + 20 (c 0.95,
methyle~zc~
chloride); Anal. Found: C, 53.38; H, 5.03; N, 23.35.
As described for the preparation of N-methyl R-1-(2-furyl)-2-aminoethanol,
2o (5R)-3-methyl-5-pyrazin-2-yl-1,3-oxazolidin-2-one (1.51 g) is treated with
1N aq.
KOH (42.1 rnL) to give R-2-( 1-hydroxy-2-N-methylamino-ethyl)-pyrazine ( 1.02
gl) as
a white solid. Mp 84 - 85 °C; Specific Rotation ~a,]D + 66 (c 0.94,
methanol).
(18) Preparation of R-2-(1-hydroxy-2-N-methylamino-ethyl)-thiazole.
As described for the preparation of N-methyl R-1-(2-furyl)-2-aminoethanol,
2-acetyl thiazole (25g) is treated with (i-Pr)3SiOTf (66.26g) and (i-Pr)ZNEt
(76.22g) in
CH2Cla (0.35L) to give 2-[1-tri-isopropylsilyloxy-vinyl]-thiazole (59.45g) as
a golden
yellow liquid. 1H-NMR (400MHz, CDCl3) S 7.80, 7.32, 5.50, 4.52, 1.35, 1.14-
1.20.
As described for the preparation of N-methyl R-1-(2-furyl)-2-aminoethanol,
2-[1-tri-isopropylsilyloxy-vinyl]-thiazole (59.45g) is treated with NCS
(29.37g) in
THF (0.35L) to give 2-[1-tri-isopropylsilyloxy-2-chloro-vinyl]-thiazole
(69.61g) as a
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yellow-orange semi-solid. 1H-NMR (400MHz, CDCl3) 8 7.77, 7.32, 6.57, 1.37,
1.10-
1.22.
As described for the preparation of N-methyl R-1-(2-furyl)-2-aminoethanol,
2-[1-tri-isopropylsilyloxy-2-chloro-vinyl]-thiazole (69.61g) is treated with
48% aq.
HF in CH3CN (0.3L) to give 2-chloroacetylthiazole (32.95g) as a light brown
liquid
which solidified to a tan solid upon cooling. 1H-NMR (400MHz, CDC13) 8 8.06,
7.79,
5.00; HRMS (ESn found 161.9779 for CSH4C1NOS+H; Anal. Found: C, 37.17; H,
2.51; N, 8.62.
As described for the preparation of N-methyl R-1-(2-furyl)-2-aminoethanol,
2-chloroacetylthiazole (24.7g) is reduced ([RuCl2(r~6 p-cymene)]2 (0.84 g),
Et3N (0.78
mL), (1R, 2R)-Np-toluenesulfonyl-1,2-diphenylethylenediamine (1.00 g) i-
propanol
(25 mL), anhydrous DMF (Aldrich Sure Seal°, 250 mL), HCOOH/Et3N (5:2,
Fluka,
55 mL) ) to give S-1-(2-thiazolyl)-2-chloroethanol (13.9 g) as a white
amorphous
solid. 1H-NMR (400MHz, CDC13) 8 7.78, 7.36, 5.24, 4.06, 3.88; % Water (KF):
0.06;
Specific Rotation [a]ZSD = 32° (c 0.83, methylene chloride); Anal
Found: C, 36.68; H,
3.58; N, 8.53; Cl, 21.37; S, 19.10; Chiral HPLC Analysis (Chiracel OJ): >
99:1.
As described for the preparation of R-2-(1-hydroxy-2-N-methylamino-ethyl)-
pyridine, S-1-(2-thiazolyl)-2-chloroethanol (6.02g) is treated with NaJ
(0.57g) and
MeNH2 (2.OM in MeOH, 0.2L) to give R-2-(1-hydroxy-2-N-methylamino-ethyl)-
2o thiazole (2.08g) as a pale yellow oil. 1H-NMR (400MHz, CDCl3) 8 7.71, 7.31,
5.68,
5.36, 3.34, 3.15, 2.64; HRMS (ESA found 159.0582 for C6H1aN20S +Hl; Specific
Rotation [a,]25D = 31° (c 1.02, DMSO); Chiral HPLC Analysis (Chiracel
OJ): >99:1.
(19) Preparation of R-2-(1-hydroxy-2-N-methylamino-ethyl)-thiophene.
As described for the preparation of N-methyl R-1-(2-furyl)-2-aminoethanol,
2-acetylthiophene (50g) is treated with (i-Pr)3SiOTf (133.6g) and (i-Pr)2NEt
(153.6g)
in CHZC12 (0.7L) to give 2-[1-tri-isopropylsilyloxy-vinyl]-thiophene (111.9g)
as a
golden yellow liquid. 1H-NMR (300MHz, CDC13) ~ 7.26, 7.22, 6.99, 4.79, 4.33,
1.32,
1.10-1.22; MS (ES+): 283.2 (M+H).
As described for the preparation of N-methyl R-1-(2-furyl)-2-aminoethanol,
2-[1-tri-isopropylsilyloxy-vinyl]=thiophene (111.19g) is treated with NCS
(58.2g) in
THF (0.6L) to afford 2-[1-tri-isopropylsilyloxy-2-chloro-vinyl]-thiophene
(140g) as a
viscous orange oil. 1H-NMR (300MHz, CDC13) 8 6.85-7.40, 5.70-5.85, 0.95-1.35.
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As described for the preparation of N-methyl R-1-(2-furyl)-2-aminoethanol,
2-[1-tri-isopropylsilyloxy-2-chloro-vinyl]-thiophene (140g) in CH3CN (0.4L) is
treated with 48% aq. HF to give 2-chloroacetylthiophene (48g) as a light brown
solid.
1H-NMR (400MHz, CDCl3) 8 7.81, 7.76, 7.19, 4.62.
As described for the preparation of N-methyl R-1-(2-furyl)-2-aminoethanol,
2-chloroacetylthiophene (26g ) is reduced ([RuCl2(r~6 p-cymene)]2 (0.99g),
Et3N (0.93
rnL), (1R, 2R)-Np-toluenesulfonyl-1,2-diphenylethylenediamine (1.18 g) i-
propanol
(25 mL), anhydrous DMF (Aldrich Sure Seal°, 250 mL), HCOOH/Et3N (5:2,
Fluka,
58 mL) ) to give S-1-(2-thienyl)-2-chloroethanol (17.8 g) as a clear colorless
liquid.
1H NMR (400 MHz, CDCl3) 8 7.30, 7.05, 7.01, 5.17, 3.76, 2.81; HRMS (En found
161.9908 for C6H7C10S; Specific Rotation [a]25D = 30 (c 0.90, methylene
chloride);
Chiral HPLC Analysis (Chiracel OJ): 99:1.
As described for the preparation of N-methyl R-1-(2-furyl)-2-aminoethanol,
S-1-(2-thienyl)-2-chloroethanol (5.54g) is treated with Et3N (l.9mL) and
methyl
isocyanate (3.36g) in CH2C12 (75mL) to give S-1-(2-thienyl)-2-chloroethanol-N-
methylcarbamate 6.87g as a clear, colorless oil. 1H NMR (400 MHz, CDC13) 8
7.30,
7.11, 7.00, 6.17, 4.77, 3.82, 2.81; HRMS (FAB) found 220.0208 for
C$HloC1N02S+Hl; % Water (KF): 0.25; Specific Rotation [a]25D = 58 (c 0.97,
methylene chloride); Anal. Found: C, 43.59; H, 4.39; N, 6.32; Chiral HPLC
Analysis
(ChiracelOJ):98.3:1.7.
As described for the preparation of N-methyl R-1-(2-furyl)-2-aminoethanol,
S-1-(2-thienyl)-2-chloroethanol-N-methylcarbamate (6.47g) in THF (35mL) is
treated
with NaH°(2.33g, 60% oil dispersion) to afford 5R-3-methyl-5-(2-
thienyl)-2-
oxazoldinone (4.94g as an ivory amorphous solid. 1H NMR (400 MHz, CDCl3) 8
7.36, 7.14, 7.01, 5.70, 3.90, 3.61, 2.95; HRMS (ESn found 184.0435 for
C8H9N02S+H; Specific Rotation [a]25D = -94 (c 1.04, methylene chloride);
(Chiracel
OJ): >99:1.
As described for the preparation of N-methyl R-1-(2-furyl)-2-aminoethanol,
5R-3-methyl-5-(2-thienyl)-2-oxazoldinone (4.63g) is treated with 1N aq. KOH
(100mL) in THF (50mL) to provide R-2-(1-hydroxy-2-N-methylamino-ethyl)-
thiophene (3.28g) as an ivory solid. 1H NMR (400 MHz, CDCl3) 8 7.26, 6.98,
4.99,
2.89, 2.48; Specific Rotation [a]25D = 26 (c 1.05, methylene chloride); Anal.
Found:
C, 53.31; H, 7.13; N, 8.84; Chiral HPLC Analysis (Chiracel OJ): >99:1.
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(20) Preparation of (R)-1-(1-benzofuran-2-yl)-2-(methylamino)ethanol.
As described for the preparation of N-methyl R-1-(2-furyl)-2-aminoethanol,
2-acetyl benzofuran (32.03g) is treated with (i-Pr)3SiOTf (80mL) and (i-
Pr)2NEt
(104mL) in CHZC12 (0.3L) to give 2-[1-tri-isopropylsilyloxy-vinyl]-benzofuran
(72.52g) as an orange oil. 1H-NMR (400MHz, CDC13) 8 7.58, 7.47, 7.30, 7.23,
5.18,
4.58, 1.23-1.42, 1.18.
As described for the preparation of N-methyl R-1-(2-furyl)-2-aminoethanol,
2-[1-tri-isopropylsilyloxy-vinyl]-benzofuran (79.52g) is treated with NCS
(29.4g) in
l0 THF (0.4L) to afford 2-[1-tri-isopropylsilyloxy-2-chloro-vinyl]-benzofuran
(79.52g)
as a viscous yellow oil.
As described for the preparation of N-methyl R-1-(2-furyl)-2-aminoethanol,
2-[1-tri-isopropylsilyloxy-2-chloro-vinyl]-benzofuran (79.52g) is treated with
48% aq.
HF (40mL) in CH3CN to give 2-chloroacetyl-benzofuran (23.32g) as a powdery,
tan
i5 solid. 1H-NMR (400MHz, CDC13) 8 7.76, 7.68, 7.61, 7.54, 7.37, 4.73; IR
(diffuse
reflectance) 2479, 2412, 2360, 2338, 2306, 1694, 1550, 1271, 1263, 1255, 1164,
1023, 752, 743, 716 cm 1; Anal. Found: C, 61.92; H, 3.68.
As described for the preparation of N-methyl R-1-(2-furyl)-2-aminoethanol,
2-chloroacetyl benzofuran (lO.Og) is reduced ([RuCla(r~6 p-cymene)]Z (0.57g),
Et3N
20 (0.47 mL), (1R, 2R)-Np-toluenesulfonyl-1,2-diphenylethylenediamine (0.56g)
i-
propanol (20 mL), anhydrous DMF (Aldrich Sure Seal°, 250 mL),
HCOOH/Et3N
(5:2, Fluka, 58 mL)) to give S-1-(2-benzofuranyl)-2-chloroethanol (9.26 .g) as
a light
brown oil. 1H NMR (400 MHz, CDCl3) S 7.58, 7.48, 7.22-7.34, 6.79, 5.10, 3.98;
IR
(liq.) 3367, 1663, 1454, 1437, 1414, 1388, 1254, 1171, 1095, 1011, 881, 808,
775,
25 753, 664 cm 1; HRMS (E~ found 196.0295 for CloH9C102; Specific Rotation
[a,]LSD =
31 (c 1.05, chloroform); Chiral HPLC Analysis (Chiracel OJ): 98:2.
As described for the preparation of N-methyl R-1-(2-furyl)-2-aminoethanol,
S-1-(2-benzofuranyl)-2-chloroethanol (8.45g) is treated with methyl isocyanate
(4.17g) and Et3N (1.74g) in CH2Cla (50mL) to give S-1-(2-benzofuranyl)-2-
3o chloroethanol-N-methylcarbamate (9.75g) as a white solid. Mp 77-
78°C; 'H NMR
(400MHz, CDC13) 8 7.58, 7.49, 7.23-7.35, 6.83, 6.11, 4.58, 3.99, 2.85; IR
(diffuse
reflectance) 3374, 1699, 1535, 1251, 1134, 975, 924, 821, 814, 771, 748, 733,
676,
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626, 613 cm 1; Specific Rotation [a]25D = 101 (c 0.85, chloroform); Anal.
Found: C,
57.04; H, 4.77; N, 5.55; Cl, 13.53.
As described for the preparation of N-methyl R-1-(2-furyl)-2-aminoethanol,
S-1-(2-benzofuranyl)-2-chloroethanol-N-methylcarbamate (9.67g) in THF (O.1L)
is
treated with KOtBu (38.5mL, 1.OM in THF) to give 5R-3-methyl-5-(2-
benzofuranyll)-
2-oxazoldinone (5.05g) as a tan solid. Mp 68-69°;1H NMR (400MHz, CDCl3)
8 7.59,
7.50, 7.35, 7.28, 6.83, 5.62, 3.87, 3.00; Specific Rotation [a]25D = -38 (c
0.95,
chloroform); Anal.Found: C, 66.14; H, 5.07; N, 6.30; chiral HPLC Analysis
(Chiracel
OJ): 96:4.
to As described for the preparation of N-methyl R-1-(2-furyl)-2-aminoethanol,
5R-3-methyl-5-(2-benzofuranyll)-2-oxazoldinone (4.89g) and 1N KOH (lOml) in
EtOH (75m1) affords 3.97g of (R)-1-(1-benzofuran-2-yl)-2-(methylamino)ethanol
as a
tan solid. Mp 88-90°C; 1H NMR (400MHz, CDC13) 8 7.56, 7.47, 7.20-7.31,
6.70,
4.92, 3.10, 3.02, 2.51; HRMS (FAB) found 192.1026 for C11Hi3NOz +Hl; Specific
15 Rotation [a]ZSD = 31° (c 1.05, chloroform); Anal. Found: C, 68.71;
H, 6.98; N, 7.16;
chiral HPLC Analysis (Chiracel OJ): 96:4.
(21) Preparation of (S)-1-(1-benzofuran-2-yl)-2-(methylamino)ethanol.
As described for the preparation of N-methyl R-1-(2-furyl)-2-aminoethanol,
20 2-chloroacetyl benzofuran (lO.Og) is reduced ([RuCl2(r~6 p-cymene)]2
(0.57g), Et3N
(0.47 mL), (1S, 2S)-Np-toluenesulfonyl-1,2-diphenylethylenediamine (0.56g), i-
propanol (20 mL), anhydrous DMF (Aldrich Sure Seal", 250 mL), HCOOH/Et3N
(5:2, Fluka, 58 mL)) to give R-1-(2-benzofuranyl)-2-chloroethanol (9.42 g) as
a light
brown oil. 1H NMR (400 MHz, CDC13) 8 7.58, 7.48, 7.22-7.34, 6.79, 5.11, 3.98;
25 HRMS (E~ found 196.0291 for CloH9C102; Specific Rotation [a]25D = -31 (c
1.03,
chloroform); Chiral HPLC Analysis (Chiracel OJ): 99:1.
As described for the preparation of N-methyl R-1-(2-furyl)-2-aminoethanol,
R-1-(2-benzofuranyl)-2-chloroethanol (8.45g) is treated with methyl isocyanate
(4.17g) and Et3N (1.74g) in CH2C12 (50mL) to give R-1-(2-benzofuranyl)-2-
3o chloroethanol-N-methylcarbamate (9.75g) as a pale brown solid. 'H NMR
(400MHz,
CDC13) 8 7.58, 7.49, 7.23-7.35, 6.83, 6.11, 4.84, 4.00, 2.85; IR (diffuse
reflectance)
3374, 1699, 1535, 1251, 1134, 975, 924, 821, 814, 771, 748, 733, 676, 626, 613
crri 1;
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Specific Rotation [a]25D = -101 (c 0.83, chloroform); Chiral HPLC Analysis
(Chiracel
OJ): 99:1.
As described for the preparation of N-methyl R-1-(2-furyl)-2-aminoethanol,
R-1-(2-benzofuranyl)-2-chloroethanol-N-methylcarbamate (10.83g) in THF (O.1L)
is
treated with KOtBu (43.1mL, 1.OM in THF) to give 5S-3-methyl-5-(2-
benzofuranyll)-
2-oxazoldinone (5.25g) as a tan solid. Mp 72-73°; 1H NMR (400MHz,
CDCl3) 8 7.60,
7.50, 7.35, 7.28, 6.87, 5.62, 3.87, 3.00; HRMS (ESA found-218.0816 for
C12H11NO3
+Hl; Specific Rotation [a]25D = 37 (c 1.00, chloroform); Anal. Found: C, C,
66.04; H,
5.13; N, 6.38; chiral HPLC Analysis (Chiracel OJ): 96:4.
As described for the preparation of N-methyl R-1-(2-furyl)-2-aminoethanol,
5S-3-methyl-5-(2-benzofuranyll)-2-oxazoldinone (S.lOg) and 1N KOH (10m1) in
EtOH (75m1) affords 4.20g of (S)-1-(1-benzofuran-2-yl)-2-(methylamino)ethanol
as a
tan solid. Mp 89°C; 1H NMR (400MHz, CDC13) 8 7.56, 7.47, 7.20-7.31,
6.70, 4.92,
3.10, 3.02, 2.51; HRMS (FAB) found 192.1026 for CilHisNOa +Hl; Specific
Rotation
[oc]25D = -30° (c 1.02, chloroform); Anal. Found: C, 68.62; H, 6.93; N,
7.25; chiral
HPLC Analysis (Chiracel OJ): 96:4.
(22) Preparation of N-(4-chlorobenzyl)-8-fluoro-1-{2-[(2-hydroxyethyl)amino]-2-
oxoethyl } -6-(morpholin-4-ylmethyl)-4-oxo-1,4-dihydroquinoline-3-carboxamide.
To a pressure tube containing methyl [3-{ [(4-chlorobenzyl)amino]carbonyl}-
8-fluoro-6-(morpholin-4-ylmethyl)-4-oxoquinolin-1(4H)-yl]acetate (4.1 g) as a
suspension in THF (40 mL) is added ethanolamine (10 mL). The mixture is
tightly
capped, heated to 60°C and stirred. After 2 days, the suspension is
cooled to room
temperature, filtered, and the precipitant washed with diethyl ether and
hexanes. The
residue is treated with acetonitrile (500 mL) and heated to reflux. The
resulting hot
suspension is quickly filtered and the filtrate cooled to room temperature and
placed in
a freezer. The resulting solid is collected, washed with diethyl ether, dried
under a
flow of air, and finally in a vacuum oven (60 °C) to yield 3.9 g of the
title compound
as a tan solid. Physical characteristics:1H NMR (d6-DMSO) 8 2.4, 3.17, 3.4,
3.6, 4.56,
4.7, 5.21, 7.4, 7.6, 8.10, 8.35, 8.77, 10.3.
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(23) Preparation of N-(4-chlorobenzyl)-1-(2-hydroxyethyl)-9-(morpholin-4-
ylmethyl)-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[1,2,3-de]quinoxaline-6-
carboxamide.
To a flame-dried flask under a nitrogen atmosphere is added N-(4-
chlorobenzyl)-8-fluoro-1-{2-[(2-hydroxyethyl)amino]-2-oxoethyl}-6-(morpholin-4-
ylmethyl)-4-oxo-1,4-dihydroquinoline-3-carboxamide (3.9 g) followed by THF (60
mL). The mixture is treated with a solution of potassium tert-butoxide in THF
(1 M,
18 mL). After 24 hours, the resulting dark red solution is diluted with
dichloromethane and partitioned against dilute pH 4 aqueous phosphate buffer.
The
l0 aqueous layer pH is adjusted to 8 and is extracted with four additional
portions of
dichloromethane. The combined organic layer is washed brine, dried over sodium
sulfate, filtered and concentrated under reduced pressure. The resulting
residue is
adsorbed onto silica and flash column chromatographed on silica eluting with
4% to
15% methanol in dichloromethane. The product containing fractions are combined
and concentrated under reduced pressure to yield 2.0 g of the title compound
as a tan
solid. Physical characteristics: 1H NMR (d6-DMSO) 8 2.4, 3.5-3.7, 4.12, 4.56,
4.92,
5.21, 7.4, 7.56, 7.84, 8.74, 10.4; MS (ESI+) for rnlz 511 (M+H)+.
(24) Preparation of N-(4-chlorobenzyl)-9-(chloromethyl)-1-(2-hydroxyethyl)-2,7-
dioxo-2,3-dihydro-1H,7H-pyrido[1,2,3-de]quinoxaline-6-carboxamide.
To a pressure tube containing N-(4-chlorobenzyl)-1-(2-hydroxyethyl)-9-
(morpholin-4-ylmethyl)-2,7-dioxo-2,3-dihydro-1H,7H-pyrido[ 1,2,3-
de]quinoxaline-6-
carboxamide (0.26g) is added 1,2-dicholroethane (10 mL) and ethyl
chloroformate
(0.5 mL). The solution is tightly capped, heated to 80°C and stirred
for 2 days. The
resulting suspension is cooled to room temperature and filtered. The collected
precipitant is washed with diethyl ether and dried in vacuo to yield 0.20 g of
the title
compound as a tan solid. Physical characteristics:1H NMR (d6-DMSO) b 3.65,
4.13,
4.57, 4.94, 5.21, 7.4, 7.69, 7.99, 8.76, 10.3; MS (ESI+) for m/z 458 (M-H).
(25) Preparation of N-(4-fluorobenzyl)-1-methyl-9-(morpholin-4-ylmethyl)-2,7-
dioxo-2,3-dihydro-1H,7H-pyrido[ 1,2,3-de]quinoxaline-6-carboxamide
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To ethyl 1-methyl-9-(morpholin-4-ylmethyl)-2,7-dioxo-2,3-dihydro-1H,7H-
pyrido[1,2,3-de]quinoxaline-6-carboxylate (1.00g) (prepared using procedures
analogous to those described in WO 02/04445) in a 25mL 1-neck round bottom
flask
equipped with a reflux air condenser and a nitrogen inlet is added 4-
fluorobenzyl
amine (0.97g) and ethylene glycol. The mixture is placed under nitrogen and is
heated
with stirring in an 80°C oil bath for 24 hours. The reaction is cooled
to room
temperature and transferred to a separatory funnel with CH2Cl2 (0.25L). The
mixture
is washed with water (0.25mL) and the organic phase is separated and dried
over
MgS04. The solid is removed by filtration, the filter cake is rinsed with
CH2Cl2
to (0.25L) and the combined filtrate is concentrated in vacuo to afford an
orange solid.
The crude material is purified on a 40S Biotage column [wet CHZCl2, eluted
CH2C12;
CH2C12/MeOH (97.5:2.5); CH2C12/MeOH (96:4); CHZCI2lMeOH (92.5:7.5);
CHZC12/MeOH (90:10)]. The product fractions are combined to provided 0.678g of
N-
(4-fluorobenzyl)-1-methyl-9-(morpholin-4-ylmethyl)-2,7-dioxo-2,3-dihydro-1
H,7H-
pyrido[1,2,3-de]quinoxaline-6-carboxamide as an ivory solid. Physical
charteristics:
mp 222-224°C; 1H-NMR (400MHz, DMSO-d6): 8 10.25, 8.73, 7.85, 7.39,
7.17, 5.21,
4.55, 3.61, 3.59; 2.40; HRMS (ESl) found 465.1938 for C25HasFNøO4 +Hl; Anal.
Found: C, 64.27; H, 5.57; N, 11.94; F, 4.08.
(26) Preparation of N-(4-fluorobenzyl)-1-methyl-9-(chloromethyl)-2,7-dioxo-2,3-
dihydro-1H,7H-pyrido[1,2,3-de]quinoxaline-6-carboxamide
To a suspension of N-(4-fluorobenzyl)-1-methyl-9-(morpholin-4-ylmethyl)-2,7-
dioxo-
2,3-dihydro-1H,7H-pyrido[1,2,3-de]quinoxaline-6-carboxamide (1.35g) in CHC13
(25mL) in a 50mL 1-neck round bottom flask equipped with a reflux condenser
and a
nitrogen inlet is added C1C02Et (0.79g) and i-Pr2NEt (0.94g). The mixture is
warmed
under reflux for 2 hours, then is cooled to room temperature and diluted with
Et20
(O.1L). The light tan solid is isolated by suction filtration, rinsed with
Et20 (O.1L), and
dried i~2 vacuo to give N-(4-fluorobenzyl)-1-methyl-9-(chloromethyl)-2,7-dioxo-
2,3-
dihydro-1H,7H-pyrido[1,2,3-de]quinoxaline-6-carboxamide (0.98g). Physical
3o characteristics:
1H-NMR (400MHz, DMSO-d6): 8 10.25, 8.75, 7.98, 7.55, 7.38, 7.16, 5.21, 4.94,
4.55,
3.93.
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Preparation 27
Heteroaryl KotBu Heteroaryl 2 Heteroaryl N,NR2
~NH2 ~~ R
OH C(=O)(Ci_4alkyl)2 O KOH/H20 OH H
27a 27b 27c
As shown in the preparation 27 above, the hetarylamino alcohol starting
material, 27a,
is converted to the oxazolidinone 27b by treatment with dimethyl carbonate (or
a Cl_
4alkyl carbonate) and a base such as potassium tart-butoxide in DMF. The
oxazolidinone, wherein R2 is Cl_4alkyl, can have the optical purity enhanced
by
crystallization or chiral chromatography. Basic aqueous hydrolysis of the
oxazolidinone with a base such as potassium hydroxide affords the optically
enhanced
l0 N-alkylated heteroarylamino alcohol 27c, wherein heteroaryl is furan and
those
hetroaryls as defined in the detailed description of the invention.
Step 1: Preparation of 5R-3-Methyl-5-(2-furyl)-2-oxazolidinone.
A 250 mL round-bottomed flask equipped with an overhead stirrer, reflux
condensor,
thermocouple and an addition funnel is charged with (R)-2-amino-1-(2-
furyl)ethanol
( 10 g) and potassium t-butoxide ( 10.6 g). Anhydrous DMF is charged at such a
rate as
to keep the temp less than 50 °C. The reaction is heated to 80
°C (internal temp), the
addition funnel is charged with dimethyl carbonate (50 mL), and the liquid is
added in
0.2 equiv portions at such a rate as to control the evolution of gas. Once
addition of
2o dimethyl carbonate is complete, the temperature is raised to reflux (about
100 °C), and
maintained for approximately 12 h. The reaction mixture is cooled to less than
60 °C,
poured into water (100 mL) and extracted with isopropyl acetate (100 mL). The
layers are separated, and the water layer is extracted with additional
isopropyl acetate
(2 x 100 mL). The combined organic layers are washed with water (100 mL) and
dried over sodium sulfate and magnesol for 10 min. The solids are removed via
vacuum filtration, and the organic layers are concentrated in vacuo. The
resulting oil
is crystallized from MTBE (2 mL/g) to provide 10.25 g of (5R)-5-(2-furyl)-3-
methyl-
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1,3-oxazolidin-2-one. Physical characteristics. 1H NMR (CDCl3, 400 MHz) 8
7.46,
6.49, 6.39, 5.47, 3.78, 2.97.
Crystallization of 5R-3-Methyl-5-(2-furyl)-2-oxazolidinone.
A 250 mL 3-neck rb flask equipped with an overhead stirrer, and thermocouple,
is
charged with 58.3 g (350 mmol, 98.4% ee) of 5R-3-Methyl-5-(2-furyl)-2-
oxazolidinone and 116.6 mL of MTBE. The resulting suspension is heated to 50
°C
(internal), resulting in the formation of a solution. The solution is allowed
to cool to
26 °C over 1.5 h, at which point a white slurry is obtained. The slurry
is cooled to 5
°C and aged for 1.5 h. The solids are collected by vacuum filtration,
rinsing with 5 °C
MTBE (1 mLlg), to give an 85% recovery (49.5g) of 5R-3-Methyl-5-(2-furyl)-2-
oxazolidinone of enriched optical and chemical purity. Physical
Charaeteristics: 1H
NMR (400 MHz, CDCl3) 8 7.46, 6.49, 6.39, 5.47, 3.78, 2.97. 13C NMR ( 100 MHz,
CDCl3, DEPT) 8 157.5, 149.8, 143.7, 110.6, 109.9, 67.5, 50.4, 31Ø Melt
Solvate:
0.04% MTBE; KF: 0.00%; Specific Rotation [a]Da5 = - 113 ° (c = 1.00,
CH2C12);
Chiral HPLC Analysis: 0.06:99.94; 99.9% ee.
Step 2: Preparation of N-Methyl R-1-(2-furyl)-2-aminoethanol [used in examples
11,23, 31].
A round-bottomed flask equipped with an overhead stirrer, reflux condensor and
nitrogen inlet is charged with (5R)-5-(2-furyl)-3-methyl-1,3-oxazolidin-2-one
(47.1 g).
A 1 M solution of KOH (987 mL) is added and the resulting solution is heated
at 50
°C. When complete, the flask is charged with NaCI (310 g) and MTBE (470
mL).
The aqueous layer is separated and further extracted twice with a solution of
MTBE
(470 mL) and CH2C12 (23 mL). The combined organic layers are dried (MgS04),
filtered, and concentrated to afford 38.0 g of the title compound. Physical
characteristics. 'H NMR (DMSO, 400 MHz) 8 7.52, 6.36, 6.24, 4.60, 2.71, 2.28;
13C
NMR (DMSO, 100 MHz) 8 157.0, 141.6, 110.1, 105.6, 65.2, 56.2, 35.9; [a,]'2D =
+
34° (EtOH, c = 1.0).
Preparation 28
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Y Het-M Heteroaryl Heteroaryl\
M = Li, Mg ~~ (Bz)Ru(TsDPEN)CI rpH
O O O O HC02H/Et3N O O
i ~
R3 R3 Rs
28a 28b 28c
1. KOtBu, THF Heteroaryl
2. R2-X ~NR2
O-
O
28d
Y = OH, MeNOMe, Ph NMe Ph " NMe , C
' N
OH
R2 = Me, Et, CH2Ph, CH2CH=CH3 R3 = t-Bu, iPr, iBu
X = CI, Br, I, OS02Ph, OS02(4-MePh), OS02CF3
As shown in the preparation 28 above, 28a is converted to 28b by treatment
with
organometallic in THF. The ketone of 28b is reduced to the chiral alcohol 28c
using a
to chiral Ruthenium catalyst and formic acid triethylamine complex.
Cyclization under
basic conditions and alkylated to give 28d, wherein hetaryl is furan and those
hetaryls
defined in the detailed description of the invention.
Step 1: Preparation of tert-Butyl (2-oxo-2-(2-furyl)-ethyl)carbamate
15 To a solution of furan (3.0 g) in anhydrous THF (30 mL) at 0 °C is
added nBuLi (2.5
M in hexane) at such a rate as to keep the internal temperature less than 25
°C. After
stirring at 0-5 °C for 50 min, the reaction mixture is cooled to
between -30 and -20
°C, and a suspension of N (tert-butoxycarbonyl)glycine N'-methoxy-N'-
methylamide
[4.36 g)] in THF (15 mL) is added drop-wise by large-bore cannula. The
resulting
2o slurry is allowed to warm to 5 °C over a 6 h period, at which time
GC indicated less
than 4% N (tert-butoxycarbonyl)glycine N'-methoxy-N'-methylamide remained. The
reaction is quenched by the addition of aqueous citric acid (20 mL; pH = 5.0
after
quench) and diluted with CHZC12 (40 mL). The resulting phases are separated,
and the
aqueous is back extracted twice with CH2Cl2 (20 mL). The organic layers are
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combined, dried over sodium sulfate, and concentrated in vacuo (50 torr) to
provide
4.1 g ( 18.2 mmol, 91 %; 91 % potency by GC) of tert-Butyl (2-oxo-2-(2-furyl)-
ethyl)carbamate 3 as a light yellow solid. Physical characteristics: 1H NMR
(CDCl3,
400 MHz), 8 = 7.60, 7.28, 6.57, 5.39, 4.50, 1.47 ppm. 13C NMR (100 MHz, CDCl3,
DEPT) 8 183.9, 155.7, 150.8, 79.9, 146.8, 117.7, 112.4, 46.9, 28.3.
Step 2: Preparation 4 of (R)-1-(2-furyl)-2-((tert-butoxycarbonyl)amino)ethanol
((r~6C6H6)Ru[(R,R)-TsDPEN]Cl ): A 50 mL flask is charged with 50 mg of
[(r~6C6H6)RuCl2]2 , 75 mg (R)(R)-TsDPEN, 2.5 mL i-PrOH (anhydrous from
Aldrich)
and 80 uL NEt3 (F.W. = 101, 0.57 mmol), a reflux condenser attached and, under
nitrogen, the reaction mixture is heated to 75°C for 1 hr. The reaction
is then cooled
to 0°C giving a dark orange solid which is collected by filtration. The
solid is washed
with diethyl ether and air-dried giving 71 mg of product (F.W. = 578.9, 0.123
mmol,
61 % as an air stable material).
To a 50 mL RB flask in the glovebox is added ca. 4 mg of (ri6C6H6)Ru[(R,R)-
TsDPEN]Cl (prepared above) followed by 5 mL of a NEt3/HCOzH (1:1.5 molar
ratio). This is allowed to stir at r.t. for 20 minutes giving a pale yellow
solution. To
this solution is added lg of tert-butyl (2-oxo-2-(2-furyl)-ethyl)carbamate and
the
reaction mixture is stirred at r.t. for 22hr (monitored by HPLC). By HPLC
analysis a
100% conversion of ketone to >99% (by area) alcohol is achieved. The crude
reaction
is poured into 75 mL H20. This is extracted 2 x 75 mL EtOAc. The combined
organic layers are extracted with 1 x 50 mL 1 M aq. NaHC03 and then with 1 x
50 mL
brine. The organic layer is dried over MgS04, filtered and concentrated to
give (R)-1-
(2-furyl)-2-((tent-butoxycarbonyl)amino)ethanol as a yellow oil, Physical
Characteristics: LCMS m/z = 250, 154; Uv-vis 7~ = 216. 1H NMR (CD3CN) ~ 7.40,
6.33, 6.25, 5.39, 4.61, 3.62, 3.30 1.36; Chiral HPLC analysis: 3: 97; 94%ee.
Step 3: preparation of 5R-3-Methyl-5-(2-furyl)-2-oxazoldinone
A round-bottomed flask equipped with an overhead stirrer, reflux condensor,
3o thermocouple and an addition funnel is charged with (R)-1-(2-furyl)-2-
((tert-
butoxycarbonyl)amino)ethanol (200 g) and anhydrous tetrahydrofuran (2.0 L). A
1.0
M solution of potassium tert-butoxide (1.06 L) is charged at such a rate as to
keep the
temp less than 35 °C. Once all of the starting material has been
consumed, as
CA 02471089 2004-06-18
WO 03/053971 PCT/US02/37613
determined by HPLC, methyl iodide (63.5 mL) is added drop-wise to the
suspension.
The suspension is then allowed to stir at 25-35 degrees until all of the
intermediate
normethyl oxazolidinone is consumed (HPLC). The reaction is quenched with
water
(2.0 L) and diluted with isopropyl acetate (2.0 L). The biphasic mixture is
agitated,
the. phases are separated, and the aqueous is back extracted with isopropyl
acetate (2 x
2.0 L). The organic layers are combined, dried over sodium sulfate (500g) and
concentrated to give 147 g of 5R-3-Methyl-5-(2-furyl)-2-oxazoldinone 28D.
Physical
characteristics. 1H NMR (CDC13, 400 MHz) 8 7.46, 6.49, 6.39, 5.47, 3.78, 2.97.
l0 Preparation 29
MeNH2
Pyres+Br3- ~ I / I (Boc)20
O~ O~gr OC O~NMe OC
IOI O O H
in situ
in situ
29a 29b 29c 29e
Preparation of tert-Butyl (2-oxo-2-(2-furyl)-ethyl)carbamate
A 250 mL RB flask is charged, in order, with; 10.56 g 2-acetylfuran, 100 mL
CHZC12
and 30.3 g pyridinium tribromide. The reaction is allowed to stir at RT for
30min
After 30min the reaction mass is cooled to 0°C and nitrogen is purged
over the surface
of the reaction mixture for 10 min to remove HBr. To the reaction mixture,
cooled to
0°C, is added 200 mL of 2 M MeNH2 (in THF) all at once. This reaction
mixture is
2o allowed to stir at 0°C for l0min after which time nitrogen is purged
through the
reaction mixture for 15 min to remove excess MeNHz. At 0°C, 42 g of
(Boc)2-O in 50
mL THF is added all at once to the reaction mixture generated above. The
reaction is
allowed to warm to RT and after 45 min an additional 5 g (Boc)2-O is added and
the
reaction mixture is stirred for an additional 30 min. The resultant reaction
mixture is
concentrated in vacuo. The residue is extracted with diethyl ether/H20. The
ether
layer is extracted with a second portion of Ha0 and then with brine. The
combined
aqueous layers are extracted with diethyl ether. The combined ether layers are
dried
over MgS04, filtered and concentrated to a purple colored oil. The residue is
dissolved in EtOAc, filtered through a plug of silica gel using 50:50 ethyl
acetate:hexane to wash the silica gel. The filtrate is concentrated giving an
oil. The
_~g_
CA 02471089 2004-06-18
WO 03/053971 PCT/US02/37613
oil is then chromatographed on silica gel using ethyl acetae/hexane (1:4)
giving 7.81
g of a white solid. Physical Characteristics: LCMS mlz = 262, 162, 140; Uv-vis
~,m~
= 200, 226, 268. 1H NMR (DMSO-d6) 8 7.98, 7.46, 6.69, 4.44 2.81, 2.78, 1.35,
1.21
ppm; 13C NMR (CDCl3) 8 184.7, 184.6; 156.3; 155.8; 146.7, 117.5; 117.3; 112.5;
113.4; 80.2; 55.4; 54.7; 35.9; 28.5; 28.3 ppm.
Preparation 30
/ I (Bz)Ru(TsDPEN)CI O / NMe
O ~~ a
HC02H/Et3N ' OHO~O
O
30a 30b
In the glovebox a fresh solution of formic acid and triethylamine is made by
the
addition of 13.2 g NEt3 (F.W. = 101, 0.13 mol, Aldrich) to 9 g 98% formic acid
(F.W. = 46, 0.196 mol, Aldrich), exothermic reaction.
To a glass vial is added 5 mg of (r~6C6H6)Ru[(R,R)-TsDPEN]Cl followed by 7
mL of the NEt3/HCOZH. This is allowed to stir at r.t. for 20 minutes giving a
pale
yellow solution. To this solution is added 1 g of the a-boc-methylaminoketone
substrate (F.W. = 239, 4.18 mmol) and the reaction mixture is stirred at r.t.
for 3 d
(monitored by HPLC). By HPLC analysis a 100 % conversion of ketone to >95% (by
area) alcohol is achieved.
The product is recovered by pouring the crude reaction into a mixture of 150
mL ethyl acetate and 150 mL H20. The aqueous layer is extracted with a second
150
mL portion of ethyl acetate. The combined organic layers are extracted with
aq.
NaHC03 (75 mL) and then brine (75 mL). The organic layer ws dried over MgSO4,
filtered and concentrated giving 1.03 g (102%) of alcohol (PHA-774326) which
is
analyzed by chiral HPLC giving 98.1 % ee of (+) isomer. 1H NMR (CD3CN) 8 7.41
(s, 1H), 6.34 (br s, 1H), 6.25 (d, J = 2.4 Hz, 1H), 4.76 (br m, 1H), 3.46 (br
overlapping, 3H), 2.74 (s, 3H), 1.37 (s, 9H).
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